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Objective To observe the effect of up-regulation of protein kinase D2 (PKD2) expression on lung ischemia-reperfusion injury (LI/RI) in mice. Methods 20 C57BL/6J wild-type (WT) mice and 40 PKD2 over-expression (PKD2+/+) mice were randomly divided into sham operation group (WS and PS groups), LI/RI group (WIR and PIR groups, the left hilum was first clamped for 1 h, then reperfused for 2 h) and the hypoxia inducible factor-1α (HIF-1α) inhibitor (2-methoxyestradiol, 2-ME) group (WIR+2ME) and (PIR+2ME) groups, 20 mg/(kg·d) 2ME was injected intraperitoneally 2 days before modeling). After 2 hours of reperfusion, the wet/dry weight ratio(W/D) of lung tissues was detected, the pathological changes of lung tissues were observed by HE staining, and the content of inflammatory factors TNF-α, IL-1β and IL-6 in lung tissues were detected by ELISA. RT-qPCR and Western blot were used to detect the expression of (HIF-1α) and vascular endothelial growth factor A(VEGFA) mRNA and protein in lung tissues. Results Compared with WS group, the W/D value of the WIR group increased, the lung tissues were damaged, the content of inflammatory factors increased, and the expression of HIF-1α and VEGFA mRNA and protein increased (P＜0.05). Compared with the WIR group, the PIR group W/D value decreased, lung tissue damage was reduced, inflammatory factor content decreased, HIF-1α and VEGFA mRNA and protein expression increased (P＜0.05). Compared with WIR and PIR groups, W/D values of (WIR+2ME) and (PIR+2ME) groups increased, lung tissue damage worsened, inflammatory factor content increased, HIF-1α and VEGFA mRNA and protein expression significantly decreased (P＜0.05). Conclusions Up-regulation of PKD2 expression can improve LI/RI, and its mechanism may be related to increasing the expression of HIF-1α/VEGFA signaling pathway.

Objectives: To investigate the protective effects of 2-methoxyestradiol (2ME) against hypoxic pulmonary hypertension (HPH) in neonatal rats., Methods: Ninety-six Wistar neonatal rats were randomly divided into a normoxia group, a hypoxia group, and a hypoxia + 2ME group, with each group further subdivided into 3-day, 7-day, 14-day, and 21-day subgroups, containing eight rats each. The hypoxia and hypoxia + 2ME groups received daily subcutaneous injections of saline and 2ME (240 μg/kg), respectively, while the normoxia group was raised in a normoxic environment with daily saline injections. Right ventricular systolic pressure (RVSP) was measured using the direct pressure method. Pulmonary vascular morphology was assessed using hematoxylin and eosin staining, with metrics including the percentage of medial thickness of small pulmonary arteries relative to the external diameter (MT%) and the cross-sectional area of the media of small pulmonary arteries relative to the total cross-sectional area (MA%). Immunohistochemistry was used to detect the expression levels of hypoxia-inducible factor-1α (HIF-1α) and proliferating cell nuclear antigen (PCNA) proteins, while real-time quantitative PCR was used to to assess HIF-1α and PCNA mRNA levels., Results: Compared to the normoxia group, the hypoxia and hypoxia + 2ME groups showed increased RVSP and upregulated HIF-1α and PCNA protein and mRNA expression levels at 3, 7, 14, and 21 days after hypoxia ( P <0.05). Furthermore, at 7, 14, and 21 days after hypoxia, the hypoxia group showed increased MT% and MA% ( P <0.05). In comparison to the hypoxia group, the hypoxia + 2ME group exhibited reduced RVSP and downregulated HIF-1α and PCNA protein and mRNA expression levels, along with decreased MT% and MA% at 7, 14, and 21 days after hypoxia ( P <0.05)., Conclusions: 2ME may protect against HPH in neonatal rats by inhibiting the expression of HIF-1α and PCNA and reducing pulmonary vascular remodeling. Citation:Chinese Journal of Contemporary Pediatrics, 2024, 26(7): 757-764 .

Objective To investigate the effect of 2-methoxyestradiol (2-ME ) on autophagy and fibrosis of dermal fibroblasts cultured under hypoxia. Methods Primary cultured human dermal fibroblasts were established, serially passaged and used for analysis at early passage (3rd to 6th generation). These fibroblasts were treated with different concentrations of 2-ME (0, 5, 10, 25 μmol/L) or the inhibitor of autophagy Bafilomycin A1 (0, 5, 10, 50 nmol/L) and cultured under hypoxia (1% O2). Then Collagen I, connective tissue growth factor (CTGF), hypoxia-inducible factor 1 alpha (HIF-1α), P62 and LC3 were detected by Western blot, and LC3 level was detected by immunofluorescence. Results Compared with those cultured in normoxia, fibroblasts cultured in hypoxia showed increased levels of HIF-1α, collagen I, CTGF, LC3II proteins and fluorescence intensity of LC3, and decreased level of P62 protein. Bafilomycin A1 decreased the levels of collagen I and CTGF and increased the levles of HIF-1α, LC3II and P62 in fibroblasts cultured in hypoxia for 24 h. Meanwhile, 2-ME inhibited the levels of HIF-1α, collagen I, CTGF and LC3II and fluorescence intensity of LC3, but improved the level of P62. These effects were in a dose-dependent manner. Conclusions 2-ME decreases fibrosis of skin fibroblasts cultivated under hypoxia probably by inhibiting autophagy. [ABSTRACT FROM AUTHOR]

Objective: To investigate the effect of 2-methoxyestradiol (2-ME2) to lymphoma Raji cells and its mechanism., Methods: Different concentrations of 2-ME2 were used to treat lymphoma Raji cells. CCK8 method was used to detect the effect of 2-ME2 to proliferation of Raji cells. Flow cytometry FITC/PI double labeling method was used to detect early apoptosis of the cells. Western blotting was used to detect the effect of 2-ME2 to the expression of BCL-2, Bax, Caspase-3 and C-myc proteins in Raji cells., Results: 2-ME2 significantly inhibited the proliferation of Raji cells. The inhibition rate increased with the increasing of drug concentration, and increased significantly with the prolongation of drug treatment time (r=0.9215). Flow cytometry FITC/PI double staining showed that the apoptotic rate of 2.5 μmol/L 2-ME2 treatment group was (33.79±1.63) %, while the apoptosis rate of the 48 h group was (51.90±2.72) %, and that of the control group was (7.08±0.36) %. After treated with 2.5 μmol/L 2-ME2 for 12 h, the expression of Bax protein was up-regulated, BCL-2 protein was down-regulated, caspase-3 protein expression was up-regulated, and C-myc protein expression was down-regulated, all of them showed a time-dependent relationship., Conclusion: 2-ME2 shows obvious inhibitory effect on lymphoma Raji cells in a dose- and time-dependent manner. Its mechanism of treatment on lymphoma Raji cells may be related to up-regulation of Bax/BCL-2 ratio and activation of Caspase-3 to induce apoptosis in cancer cells. Down-regulation of C-myc protein expression also participates in the apoptotic process.

Objective: To explore the signal pathway that mediates the effect of 2-methoxyestradiol（2ME2） on human laryngeal papilloma cell line, in terms of cell proliferation and neovascularization. Method: HIF-1α expression of human laryngeal papilloma cell line（Hs840. T） was interfered using siRNA, and the cells were then processed by 2ME2 in two concentrations. RT-PCR and ELISA were performed to detect the difference of HIF-1α in cells with normal or lower HIF-1α mRNA level, with ELISA test of excretory VEGF level and CCK8 test of cell viability. Result: The IC50of 2ME2 in Hs840. T was 0.309 μmol/L in terms of the inhibition effect of cell proliferation（ P <0.01）. Baseline level of intracellular HIF-1α was detectable, and procession of Hs840. T cells by 2ME2 of 0.4 μmol/L inhibited the transcription and expression of HIF-1α by（76.8±2.0）% and（68.6±3.5）% [vs blank group（100.0±2.7）% and（100.0±6.9）%, P <0.01]. VEGF excretion decreased to（50.8±2.1） and（28.1±4.0）% [vs blank group（100.0±3.1）%, P <0.01]after procession by 2ME2 of 0.2 μmol/L and 0.4 μmol/L. After the successful interference of HIF-1α by siRNA, the inhibition effect on cell proliferation by 2ME2 of 0.4 μmol/L decreased to（51.5±3.8）% [vs control group（65.7±1.7）%, P <0.01]. siRNA interference of HIF-1α lead to a decrease of HIF-α mRNA and protein level to（16.3±0.9）% and（7.4±0.8）% [vs cells not interfered（76.8±2.0）% vs（68.6±3.5）%, P <0.01]. Secretory VEGF dropped to（41.0±2.9）% [vs cells not interfered（28.1±4.0）%, P <0.05]. Conclusion: 2ME2 has a significant inhibitory effect on human laryngeal cell line. The inhibition of cell proliferation was mediated by a lower level of HIF-1α and therefore lower VEGF. 2ME2 might serve as a novel potential therapy for patients of recurrent respiratory papillomatosis., Competing Interests: The authors of this article and the planning committee members and staff have no relevant financial relationships with commercial interests to disclose., (Copyright© by the Editorial Department of Journal of Clinical Otorhinolaryngology Head and Neck Surgery.)

Objective: To detect the effects of quercetin (Que) combined with 2-methoxyestradiol (2-ME) on the proliferation of androgen-dependent LNCaP human prostate cancer cells line and androgen-independent PC-3 human prostate cancer cells line, and to evaluate the antitumor effects of different combos of the two drugs., Methods: After LNCaP and PC-3 cells were treated with different concentration of quercetin (0, 3.125, 6.25, 12.5, 25, 50, 100, 200 μmol/L) or 2-ME (0, 0.312 5, 0.625, 1.25, 2.5, 5, 10 μmol/L) for 48 h, the inhibitory rates of cell growth were tested using trypan blue staining method respectively. Then the concentration-effect curves were drawn and IC(50) values were calculated. According to the fitted dose-effect curves and IC(50) values, appropriate concentrations of quercetin and 2-ME were selected to compose 16 different combos. Then cells were treated with different combos of Que and 2-ME for 48 h, and then the growth inhibitory rates of cell growth were detected. According to the equation and median-effect principle, the CI values of 16 different combos of Que and 2-ME were calculated to evaluate their antitumor effects., Results: The inhibition rate of LNCaP or PC-3 cell growth treated with varying doses of quercetin or 2-ME alone showed a dose-dependent increase respectively. The IC(50) values of quercetin and 2-ME were 23.29 μmol/L and 1.89 μmol/L for LNCaP cells; and 22.12 μmol/L and 1.74 μmol/L for PC-3 cells respectively. After treated with 16 combos of Que (5, 10, 20, 40 μmol/L) and 2-ME (0.5, 1, 3, 5 μmol/L) for 48 h, for LNCaP cells, lower dose of Que (5 and 10 μmol/L) with higher dose of 2-ME (3 and 5 μmol/L) showed synergistic activity, whereas for PC-3 cells, besides the above combination of Que 10 μmol/L and 2-ME 3 μmol/L, higher dose of quercetin (20 and 40 μmol/L) with higher dose of 2-ME (3 and 5 μmol/L) also showed synergistic activity., Conclusions: Both quercetin and 2-methoxyestradiol could inhibit the growth of LNCaP and PC-3 human prostate cancer cells in a dose dependent manner. We confirmed that combinations of quercetin and 2-ME at appropriate concentrations have the potential for synergetic antiproliferative activity in vitro.

Objective: To explore the effect of 2-methoxyestradiol (2-ME2) on apoptosis of human acute T lymphoblastic leukemia cells, and its underlying mechanism., Methods: The growth inhibition of CEM cells was detected by MTT assay; apoptotic cells were detected by DNA laddering analysis; the expressions of P53 mRNA and protein were detected by RT-PCR and Western blot respectively., Results: 2-ME2 remarkably inhibited the CEM cell growth and the 50% growth inhibitory concentration (IC50) at 48 h was 2 µmol/L. The DNA ladder could be detected in CEM cells after treating with 2 µmol/L 2-ME2 for 24, 48 and 72 hours; after treating with 2 µmol/L 2-ME2 for 24, 48 and 72 hours, a time-dependent reduction of P53 mRNA and protein expressions was found in CEM cells., Conclusion: The anti-leukemia effect of 2-ME2 is completed through the induction of cell apoptosis. Down-regulation of P53 gene expression may be an underlying mechanism.

This study was aimed to explore the synergistic effect of 2-methoxyestradiol (2-ME2) and bortezomib (Bor) on the proliferative inhibition and apoptosis of U266 cell line and its possible mechanism. The cells were treated with 2-ME2, Bor alone and 2-ME2 combined with Bor, respectively. The cell viability and proliferative curve were detected by CCK8, the cell apoptosis was detected by caspase 3/7 activity test, cell cycle status was analyzed by flow cytometry, and real-time PCR was used to detect the mRNA expression of P21, BAX and BCL-2. The results showed that compared with cells treated with 2-ME2 or Bor alone, the proliferative potential of cells in combination group was significantly inhibited (p < 0.05), and apoptosis rate markedly increased (p < 0.05), cell cycle was arrested at G(1)-S phase, the mRNA expressive level of P21 and BAX increased, while the expression of BCL-2 decreased. It is concluded that 2-ME2 combined with Bor synergistically inhibits cell proliferation and induces apoptosis in U266 cell line. The possible mechanism may be associated with its effect of up-regulating P21 and BAX expressions.

The purpose of this study was to explore the mechanism underlying the regulation of 2-methoxyestradiol (2-ME)-induced cell apoptosis by mcl-1 and bax gene in myelodysplastic syndrome (MDS). The MUTZ-1 cells were pretreated with 2-ME; then the activity of caspases-3 was determined by fluorescent colorimetry; the mRNA expressions of apoptosis-related genes (mcl-1) and bcl-2-related X protein (bax) were determined by RT-PCR. The results showed that as compared with control, the 2-ME enhanced the activity of caspase-3 in MUTZ-1 cells in a dose-and time-dependent manners (p<0.05); along with increasing of 2-ME concentration, the expression of intracellular mcl-1 mRNA reduced (p<0.05), meanwhile the expression level of mcl-1 mRNA negatively correlated to the activity of caspase-3 at the corresponding time points (r=-0.992, p<0.01), but the expression of bax mRNA did not show significant change (p>0.05). It is concluded that 2-ME can regulate the apoptosis of MDS cells through the pathway of down-regulating the expression of mcl-1 mRNA and activating the caspase-3.

The aim of this study was to investigate apoptotic effect of 2-methoxyestradiol (2-ME2) on K562 cells and its mechanism. K562 cells were treated with different concentrations of 2-ME2. MTT assay was used to examine the effect inducing growth inhibition. DNA fragmentation assay and Annexin V-FITC/PI staining were used to detect the effect of apoptosis. The change of mitochondrial transmembrane potential was analyzed by flow cytometry. The expressions of related gene mRNA and/or proteins were detected by RT-PCR and Western blot respectively. The results indicated that the 2-ME2 inhibited proliferation of K562 cells in a time- and dose-dependent manners and the concentration of 50% growth inhibition (IC(50)) was 2 micromol/L at 48 hours. 2-ME2 induced DNA ladder and significantly increased apoptosis in K562 cells when exposed to 2 micromol/L of 2-ME2 for 24, 48 and 72 hours, the result of Annexin-V/PI staining showed that rates of the apoptotic cells were 13.78%, 22.32% and 29.43% respectively, which was remarkably higher than that of control (1.78%) (p < 0.05). The FCM analysis showed that the mitochondrial transmembrane potential in K562 cells lowered after exposed to 1, 2 and 4 micromol/L of 2-ME2 for 24 hours. 2-ME2 down-regulated the expression of bcr/abl and bcl-2, up-regulated the expression of bax mRNA, and down-regulated protein expressions of bcl-2, procaspase-3, procaspase-9, PARP (116 kD) and p-Akt, and up-regulated expression of cytoplasmic Cyto-C and PARP 85 kD apoptosis-related cleavage fragment protein, but had no effect on total Akt protein in K562 cells after treated with 2 micromol/L of 2-ME2 for 24, 48 and 72 hours. It is concluded that the 2-ME2 can induce the apoptosis and inhibit the proliferation of K562 cells by increasing the ratio of bax/bcl-2, reducing the mitochondrial transmembrane potential, releasing cytochrome C to cytoplasm, initiating the mitochondrial apoptosis pathway and leading in turn to caspase-3 activation. These findings suggest that interfere PI3K/Akt signal pathway via down-regulating the expression of bcr/abl mRNA is implicated in the effect of 2-ME2 on K562 cells.

The aim of this study was to investigate the effects of 2-methoxyestradiol (2-ME) on expressions of caspase-3 and survivin in chronic myelocytic leukemia (CML) K562 cells. The experiment was divided into 3 groups: control group, in which K562 cells were cultured in medium without 2-ME; the experimental group, in which K562 cells were cultured in medium containing different concentrations of 2-ME (1, 2, 4, 8 and 16 micromol/L) for 36 hours; the negative control group, in which K562 cells were replaced by distilled water without RNase in medium. The apoptosis rate, the protein and its mRNA expressions of caspase-3 and survivin of K562 cells was detected by TUNEL, flow cytometry (FCM), half-quantitative RT-PCR respectively. The results showed that the apoptosis rate of K562 cells in experimental group was significantly higher than that in control group (p < 0.05). The apoptosis rate of K562 cells detected by FCM was almost the same as that detected by TUNEL method (p < 0.01). The result detected by TUNEL methods was positively correlated with that detected by FCM (gamma = 0.845, p = 0.034). The expression of caspase-3 protein increased in a concentration-dependent manner, and also this expression level in the experimental group was higher than that in the control group (p < 0.05); the expression of survivin protein decreased along with the increasing of 2-ME concentration. and the difference between the experimental group and the control group was statistically significant (p < 0.05). The expression of caspase-3 mRNA was higher in the experimental group than that in the control group (p < 0.01), and the expression of survivin mRNA was lower in the experimental group than that in the control group (p < 0.01). The expression level of caspase-3 mRNA was negatively correlated with that of survivin (gamma = -0.966, p = 0.001). It is concluded that the 2-ME can induce apoptosis of K562 cells in a concentration-dependent manner and indicate its promising potential in the treatment of CML patients.

Objective: To investigate the effects of low-dose 2-methoxyestradiol (2ME2) on the expression of two isoforms of PRDM1 gene (PRDM1alpha with and PRDM1beta without PR-domain) during the myeloma cell differentiation., Methods: In myeloma cell line NCI-H929, RPMI8226, KM3 and LP-1, PRDMla and PRDM1beta transcripts were detected by real-time quantitative PCR after treatment with low-dose of 2ME2 (0.5 micromol/L) for 0 h, 48 h and 72 h., Results: Both PRDM1alpha and PRDMIbeta3 were time-dependently upregulated, and the PRDMlalpha/beta ratio was elevated from 1.461 +/- 0.033 to 2.663 +/- 0.381 (P < 0.01), 1.929 +/- 0.334 to 2.727 +/- 0.362 (P < 0.05), 1.471 +/- 0.012 to 4.367 +/- 0.243 (P < 0.01) and 1.660 +/- 0.042 to 3.059 +/- 0.167 (P < 0.01) in NCI-H929, RPMI8226, KM3 and LP-1 cells respectively., Conclusion: Low dose of 2ME2 can induce differentiation of myeloma cells as well as up-regulate the expression of PRDMlalpha and PRDM1beta in these cells. Elevation of PRDM1alpha/beta ratio was in a time-dependent manner and positively associated with cell differentiation and in accordance with Ying-Yang mechanism of PR-domain-containing gene family.

Objective: To investigate the repression of c-myc induced by Blimp-1 in 2-methoxyestradiol(2ME2)-mediated differentiation of multiple myeloma cells., Methods: CZ-1 and LP-1 myeloma cells lines were exposed to 0.5 micromol/L of 2ME2 and 0.5 micromol/L of 2ME2 + antisense oligonucleotides (ASODN) for 72 h. The effects on transcription of c-myc mRNA were studied by RT-PCR. The c-myc protein was assayed with Western blot. The changes of the cell lines in morphology, expression of surface CD49e and quantity of immunoglobin light chain secretion in the supernatant were studied., Results: Incubation of the cells with 0.5 micromol/L of 2ME2 could up-regulated Blimp-1 expression and increase the expression of c-myc gene. In contrast, c-myc expression was decreased with Blimp-1 expression down-regulated by ASODN and the cell lines differentiation was arrested., Conclusion: Blimp-1 could directly repress the expression of c-myc in 2ME2-mediated differentiation induction of multiple myeloma cells.

Objective: To study the effect of 2-methoxyestradiol(2-ME) on telomerase activity expression and apoptosis in human myelodysplastic syndrome cells line MUTZ-1., Methods: MUTZ-1 cells were incubated with different concentrations of 2-ME, apoptosis rate and cell cycle were measured by flow cytometry (FCM). Telomerase activity in MUTZ-1 cells was examined by telomeric repeat amplification protocol-Enzyme linked immunosorbent assay (TRAP-ELISA)., Results: The FCM analysis showed that cells in G0/G1 phase and S phase were decreased, while in G2/M phase increased after exposed to 1,2 and 4 micromol/L of 2-ME for 12 hours (P < 0.05). 1 and 2 micromol/L of 2-ME had no notable effect on MUTZ-1 cells as compared with the control group (P > 0. 05). Cells incubated with 1, 2 and 4 micromol/L of 2-ME for 36 hours were induced apoptosis, the percentage of apoptosis was between (12.87 +/- 0.86)% and (21.82 +/- 1.71)% with a dose- and time- dependent manner. Telomerase activity was significantly inhibited in these concentration and negatively correlated with cell number in G2/M phase (r = -0.979, P = 0.021) and increased apoptosis (r = -0.970, P = 0.030 ), respectively. Moreover, the inhibition effect of telomerase activity was enhanced in a dose- and time- dependent manner., Conclusions: 2-ME-induced apoptosis and inhibition of telomerase activity provide a possible mechanism for explaining the 2-ME's anticancer activity.

In order to investigate the differentiation-inducing effect of 2-methoxyestradiol (2ME2), an estrogen derivative, on CD138+ (Syndecan-1) primary myeloma cells from 7 patients with myeloma, primary myeloma cells from 7 patients were enriched by using CD138 immunomagenetic beads. The enriched CD138+ (Syndecan-1) myeloma cells treated with 0.5 micromol/L 2ME2 for 36 h and 72 hours were used to observe the differentiation changes with expression of CD49e and to quantitatively detect the light-chain secretion in the supernatants. The results indicated that 2ME2 caused morphological and immunophenotypic changes with light-chain secretion in the supernatant, and the typical features of differentiation of CD138+ (Syndecan-1) primary myeloma cells. CD138+ (Syndecan-1) myeloma cells became more and more mature in morphology, with decreased ratio of nucleus to plasma, disappearance of nucleoli, and pyknosis of chromatin. The result of detecting the expression of CD49e of CD138+ (Syndecan-1) myeloma cells showed the obvious up-regulation of CD49e expression after exposure to 2ME2 for 72 hours. Quantitative assay for light chain secretion in the supernatant of bone marrow CD138+ cells from patients showed remarkable increment at 72 hours. It is concluded that lower concentrations of 2-methoxyestradiol can induce differentiation of bone marrow CD138+ cells from multiple myeloma patients.

Objective: To evaluate the effects of 2-methoxyestradiol (2-ME(2)) on human cervical cancer HeLaS3 cells and cervical cancer xenografts., Methods: Cell proliferation assay and cell cycle analysis were used to measure HeLaS3 cell growth and cell cycle progression after 2-ME(2) treatment. Fluorescent microscopy to observe the cell morphology and DNA electrophoresis to measure apoptosis. In addition, the effect of 2-ME(2) on the expression of inducible nitric oxide synthase (iNOS) was measured by Western blot. Moreover, human cervical cancer model was set up using HeLaS3 cells and 2-ME(2) (75 mg/kg) was orally given for 14 d. Tumor volume was determined and apoptosis was detected by in situ cell death., Results: Newly-formed cell amount in treated group was 81% of that in control group after 1 micromol/L 2-ME(2) treatment for 48 h (P < 0.05), and was 19% of that in control group after 2-ME(2) treatment for 96 h (P < 0.01). G(2)/M phase cells were increased to 55% from 16% of the control (P < 0.01), and apoptotic cells were increased to 16% from 4% of the control, after 5 micromol/L 2-ME(2) treatment for 20 h. Nuclear condensation and abnormal metaphase cells were found by fluorescent microscopy. Typical DNA ladder was found by DNA electrophoresis. And the expression of iNOS was increased by 2-ME(2) in a time- and concentration-dependent manner, in parallel with apoptosis. Moreover, apoptosis was prevented by the iNOS inhibitor 1400W. In vivo, tumor volume was reduced 34% while compared with the control group. In situ cell death detection found more apoptotic and necrotic cells in 2-ME(2)-treated group., Conclusions: 2-ME(2) inhibits human cervical cancer HeLaS3 cells and tumor growth in cervical cancer xenografts. Thus 2-ME(2) has the therapeutic potential for cervical carcinoma.

Objective: To explore the effect of 2-methoxyestradiol (2ME2) and arsenic trioxide (As2O3) on the apoptosis related gene expression in multiple myeloma cell line CZ-1., Methods: Total RNA was isolated from CZ-1 cells which had been treated with 2ME2 and As2O3 at an apoptosis-inducing concentration and reverse-transcribed into a cDNA probe labeled with Bio-16-dUTP, and then hybridized it with a microarray containing up to 96 key genes involved in apoptosis. The gene expression profile of the 2ME2 and As2O3 treated CZ-1 cells were analyzed with GEArray Analyzer software. The microarray results were confirmed by RT-PCR., Results: As2O3 treatment caused the alteration in the expression of 52 genes (54.2% of total genes on microarray). Among them, 42 (80.8%) were upregulated and 10 (19.2%) were downregulated. The upregulated genes were mainly involved in caspases family, P53 and ATM pathway, death effector domain family, TNF receptor family and CIDE family. 2ME2 treatment resulted in the alteration of 42 genes (43.8% of the total genes on microarray). Of them, 32 (76.2%) were downregulated and 10 (23.8%) were upregulated. The downregulated genes mainly belonged to bcl-2 family, inhibitor of apoptosis protein family (IAP), TRAF family, TNF ligand family, and CARD family., Conclusion: 2ME2 and As2O3 induce the CZ-1 cells apoptosis by different pathways. As2O3 mainly induces upregulation of proapoptotic genes, and 2ME2 downregulation of anti-apoptotic genes expression.

The objective was to explore the in vitro effects of growth inhibition and apoptosis induction of 2-methoxyestradiol (2ME2), an estrogen derivative, on seven myeloma cell lines NCI-H929, HS-sultan, KM3, SKO-007, CZ-1, U266 and LP-1and to observe its synergistic effects in combination with some other drugs, such as dexamethasone, As(2)O(3), thalidomide and zoledronic acid. Seven myeloma cell lines NCI-H929, HS-sultan, KM3, SKO-007, CZ-1, U266 and LP-1 were cultured at different concentrations with or without dexamethasone, As(2)O(3), thalidomide and zoledronic acid. Cell viability was assessed by trypan blue assay, plasma cell labeling index (PCLI) was detected by BrdU assay, terminal-deoxynucleotidyl transferase mediated dUTP nick end labeling (TUNEL) assay were used to determine apoptosis cells in situ. Synergistic effects of 2ME2 in combination with other drugs were judged by King's formula. The results showed that after treatment with 1, 4, 8, 12, 16 micromol/L 2ME2 at 12, 24, 36 and 48 hours respectively, 2ME2 caused a dose- and time-dependent inhibition of the cell viability. The concentration of 50% growth inhibition (IC(50)) was between (20.8 +/- 0.27) and (34.1 +/- 0.57) micromol/L. After treatment with 12 micromol/L 2ME2 within 24 hours, 2ME2 led to a progressive decline in the fraction of S-phase cells by BrdU assay, plasma cell labeling index (PCLI) declined from (30.14 +/- 4.28)% to (14.71 +/- 6.27)% (P < 0.05). After treatment with 1, 4, 8, 12, 16 micromol/L 2ME2 at 12, 24, 36 and 48 hours respectively, 2ME2 can induce a dose- and time-dependent apoptosis of myeloma cell lines. The percentage of apoptosis was between 9% - 33% (P < 0.05). Q value of synergistic effects was between 1.13 to 1.43. It is concluded that 2ME2 can inhibit proliferation and induce apoptosis of myeloma cell lines and has synergistic effects with dexamethasone, As(2)O(3), thalidomide and zoledronic acid.

Objective: To investigate the differentiation induction effect of 2-methoxyestradiol (2ME2), an estrogen derivative on myeloma cell line CZ-1., Methods: The changes of CZ-1 cells in morphology, expression of surface CD49e and quantity of light chain secretion in the supernatant were observed when treated with 0.1 approximately 0.5 micromol/L 2ME2 for 48 h., Results: 2ME2 could induce differentiation of CZ-1 cells. The cells appeared decreased in size of nucleus, increased in cytoplasma, decreased in the ratio of nucleus to plasma, decreased in number or disappearance of nucleolus, and thickness and pyknosis of chromatin. The expression of CD49e was increased from (12.20 +/- 1.57)% to (24.80 +/- 1.26)% (P < 0.05). Light chain secretion in the supernatant was increased from (35.97 +/- 2.60) microg/ml to (79.67 +/- 1.88) microg/ml (P < 0.05)., Conclusion: Low concentrations of 2ME2 could induce differentiation of myeloma cell line CZ-1.