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Background and purpose: Leptomeningeal metastasis is a form of central nervous system metastasis of melanoma. High mobility group A2 (HMGA2) has been proven to play an important role in the occurrence and development of various tumors, but its biological functions in leptomeningeal metastatic melanoma cells remain unclear. On the basis of building mouse models of central nervous system metastasis of melanoma, this study investigated the differences in cell migration and cell proliferation among leptomeningeal metastatic melanoma cells, primary site melanoma cells and brain parenchymal metastatic melanoma cells, and further clarified the effects of differentially expressed gene HMGA2 on cell migration and proliferation of leptomeningeal metastatic melanoma cells. Methods: B16 mouse melanoma cells (B16-parental cells, B16-Par) stably expressing tdTomato and luciferase were generated by lentiviral infection. Subsequently, B16 specific brain parenchymal metastatic cells (B16-brain metastatic cells, B16-BrM) and B16 specific leptomeningeal metastatic cells (B16-leptomeningeal metastatic cells, B16-LM) were collected after adaptive screening of metastatic sites in vivo. The differences in migration and proliferation among B16-Par, B16-BrM and B16-LM were assessed by wound healing assay and cell counting kit-8 (CCK-8). RNA sequencing (RNA-seq) was used to analyze differential gene expression in B16-Par, B16-BrM and B16-LM, and HMGA2 gene specifically upregulated in B16-LM was screened out. The results were verified by real-time fluorescence quantitative polymerase chain reaction (RTFQ-PCR) and Western blot. Gene ontology (GO) analysis was performed for genes which were upregulated in B16-LM specifically. siRNA was used to interfere with the expression of HMGA2 gene in B16-LM, and the knock-down effect was verified by RTFQ-PCR and Western blot. The effects of knocking down HMGA2 on cell migration and proliferation were detected by wound healing assay and CCK-8 assay. Using GSE174401 data in Gene Expression Omnibus (GEO), the specificity of HMGA2 gene expression in leptomeningeal metastatic melanoma cells from patients was verified. Results: Compared with Par cells, tumor cells screened by the brain environment were more likely to colonize the central nervous system. B16-LM had stronger migration and proliferation abilities, and upregulated the expression of HMGA2 gene. GO analysis revealed that HMGA2 was associated with many biological processes such as angiogenesis and cell proliferation. When the expression of HMGA2 gene was knocked down, the migration and proliferation of B16-LM could be inhibited. HMGA2 was upregulated in leptomeningeal metastatic melanoma cells from patients. Conclusion: Leptomeningeal metastatic melanoma cells had relatively unique cellular characteristics, which promoted cell migration and proliferation by upregulating HMGA2 gene expression.

Background and purpose: Leptomeningeal metastasis is a form of central nervous system metastasis of melanoma. High mobility group A2 (HMGA2) has been proven to play an important role in the occurrence and development of various tumors, but its biological functions in leptomeningeal metastatic melanoma cells remain unclear. On the basis of building mouse models of central nervous system metastasis of melanoma, this study investigated the differences in cell migration and cell proliferation among leptomeningeal metastatic melanoma cells, primary site melanoma cells and brain parenchymal metastatic melanoma cells, and further clarified the effects of differentially expressed gene HMGA2 on cell migration and proliferation of leptomeningeal metastatic melanoma cells. Methods: B16 mouse melanoma cells (B16-parental cells, B16-Par) stably expressing tdTomato and luciferase were generated by lentiviral infection. Subsequently, B16 specific brain parenchymal metastatic cells (B16-brain metastatic cells, B16-BrM) and B16 specific leptomeningeal metastatic cells (B16-leptomeningeal metastatic cells, B16-LM) were collected after adaptive screening of metastatic sites in vivo. The differences in migration and proliferation among B16-Par, B16-BrM and B16-LM were assessed by wound healing assay and cell counting kit-8 (CCK-8). RNA sequencing (RNA-seq) was used to analyze differential gene expression in B16-Par, B16-BrM and B16-LM, and HMGA2 gene specifically upregulated in B16-LM was screened out. The results were verified by real-time fluorescence quantitative polymerase chain reaction (RTFQ-PCR) and Western blot. Gene ontology (GO) analysis was performed for genes which were upregulated in B16-LM specifically. siRNA was used to interfere with the expression of HMGA2 gene in B16-LM, and the knock-down effect was verified by RTFQ-PCR and Western blot. The effects of knocking down HMGA2 on cell migration and proliferation were detected by wound healing assay and CCK-8 assay. Using GSE174401 data in Gene Expression Omnibus (GEO), the specificity of HMGA2 gene expression in leptomeningeal metastatic melanoma cells from patients was verified. Results: Compared with Par cells, tumor cells screened by the brain environment were more likely to colonize the central nervous system. B16-LM had stronger migration and proliferation abilities, and upregulated the expression of HMGA2 gene. GO analysis revealed that HMGA2 was associated with many biological processes such as angiogenesis and cell proliferation. When the expression of HMGA2 gene was knocked down, the migration and proliferation of B16-LM could be inhibited. HMGA2 was upregulated in leptomeningeal metastatic melanoma cells from patients. Conclusion: Leptomeningeal metastatic melanoma cells had relatively unique cellular characteristics, which promoted cell migration and proliferation by upregulating HMGA2 gene expression. [ABSTRACT FROM AUTHOR]

Objective: To investigate the expression of peroxiredoxin 1 (PRDX1), high mobility group AT-Hook 2 (HMGA2) and formin like 2 (FMNL2) in gastric cancer tissues and the relationship with clinicopathological features, epithelial-mesenchymal transition (EMT) and prognosis. Methods: A total of 153 gastric cancer patients who were admitted to our Hospital from January 2017 to February2018 were selected, and intraoperative cancer tissues and adjacent tissues were collected. PRDX1, HMGA2, FMNL2, vimentin (VIM),epithelial-cadherin (E-cad) positive expression were detected by immunohistochemistry, and PRDX1, HMGA2, FMNL2, VIM, E-cad relative expression amount were detected by real time quantitative PCR (qRT-PCR) method. The relationship between PRDX1, HMGA2and FMNL2 expression in gastric cancer tissues and their clinicopathological features, EMT and prognosis were analyzed. Results:Compared with adjacent tissues, the positive expression rate and relative expression amount of PRDX1, HMGA2, FMNL2 and VIM in gastric cancer tissues increased, while the positive expression rate and relative expression amount of E-cad decreased (P<0.05). The positive expression rates of PRDX1, HMGA2 and FMNL2 in gastric cancer tissues were related to TNM stage, lymph node metastasis and distant metastasis (P <0.05). Pearson correlation analysis showed that the relative expression amount of PRDX1, HMGA2 and FMNL2 mRNA in gastric cancer tissues were positively correlated with the relative expression amount of VIM mRNA (r=0.562, 0.517,0.621, all P<0.05), and negatively correlated with the relative expression amount of E-cad mRNA (r=-0.603,-0.544,-0.574, all P<0.05). The 3-year cumulative survival rate of 153 gastric cancer patients was 68.44% (106/153). Kaplan-Meier survival curve analysis showed that the 3-year cumulative survival rate of PRDX1, HMGA2 and FMNL2 positive group were lower than those of negative group (P<0.05). Conclusion: The expression of PRDX1, HMGA2 and FMNL2 increased in gastric cancer tissues, which is related to TNM stage, lymph node metastasis, distant metastasis, EMT and prognosis. It can be used as an auxiliary evaluation index for the condition and prognosis of gastric cancer. [ABSTRACT FROM AUTHOR]

Objective To investigate how miRNA let-7a targets high-mobility AT-hook 2 (HMGA2) and suppresses the proliferation of laryngeal carcinoma cell line TU212. Methods let-7a mimics was synthesized and transiently transfected into laryngeal carcinoma cell line TU212 by cationic liposome. TU212 cells were injected subcutaneously into nude mice to construct xenograft. The expression of let-7a and HMGA2 in TU212 cells and xenograft in the nude mice were detected by RT-qPCR and Western blot. Results In laryngeal carcinoma TU212 cells with let-7a overexpression, the expression of HMGA2 at mRNA and protein levels were down-regulated, and cell proliferation ability was inhibited. Compared with NC group and blank control group, the weight and volume of laryngeal carcinoma cell TU212 transfected with let-7a mimics were significantly decreased in vitro by tumorigenesis test; the expression of HMGA2 at mRNA and protein levels were significantly down-regulated. Conclusion let-7a could significantly inhibit the expression of HMGA2 mRNA and protein, and further inhibit the proliferation of laryngeal cancer cells.

BACKGROUND: MicroRNA-98-5p (miR-98-5p) can inhibit the high mobility group AT-HOOK 2 (HMGA2). The phosphatidylinositol 3-kinase/alkaline phosphatase/glycogen synthase kinase-3β (PI3K/AKT/GSK-3β) signaling pathway is involved in cell proliferation. During the fracture healing process, it is unclear whether miR-98-5p can act on the PI3K/Akt/GSK-3β pathway by activating HMGA2 to promote the proliferation of osteoblasts. OBJECTIVE: To investigate the mechanisms of microRNA-98-5p (miR-98-5p) in promoting osteoblast proliferation and differentiation by regulating HMGA2 and PI3K/Akt/GSK-3β pathway. METHODS: MC3T3-E1 cells transfected with miR-98-5p mimics and HMGA2 plasmid by Lipofectamine 2000 were divided into a miR-98-5p mimics group and a HMGA2 overexpression group, respectively. MC3T3-E1 cells transfected with dimethyl sulfoxide and scrambled plasmids were divided into a mimic control group and a scramble group. Cells transiently transfected with miR-98-5p inhibitor using liposomes were as a miR-98-5p inhibitor group. Normal MC3T3-E1 cells without treatment were used as a blank control group. RESULTS AND CONCLUSION: There were no significant differences in HMGA2, PI3K/Akt/GSK-3β, alkaline phosphatase activity and mRNA level between the blank control group and the mimic control group. Compared with the mimic control group, the HMGA2 and PI3K/Akt/GSK-3β protein expression, cell differentiation and proliferation, alkaline phosphatase activity and mRNA level were significantly reduced in the miR-98-5p mimics group. The interaction between miR-98-5p and HMGA2 predicted by Targetscan7.1 showed that compared with the mimic control group, the HMGA2 protein and mRNA were reduced in the miR-98-5p mimic group, while compared with the miR-98-5p mimic group, the HMGA2 protein and mRNA had an increase in the miR-98-5p inhibitor group. There were no significant differences in osteoblast proliferation, alkaline phosphatase activity and mRNA between the blank control group and the scramble group. Compared with the scramble group, the PI3K/Akt/GSK-3β expression, cell differentiation and proliferation, alkaline phosphatase activity and mRNA in the HMGA2 overexpression group were significantly increased. To conclude, miR-98-5p can inhibit the HMGA2 and PI3K/Akt/GSK-3β expressions in MC3T3-E1 cells, inhibit cell proliferation and differentiation. HMGA2 is possible the direct target of miR-98-5p. [ABSTRACT FROM AUTHOR]

Objective To elucidate the role of miR-490-3p in regulating HMGA2 expression and affecting epithelial-mesenchymal transition (EMT) of SW1990 cells. Methods miR-490-3p expression was measured in HPDE and SW1990 cell lines by Real-time PCR. SW1990 cell lines were cultured in vitro and divided into four groups, including inhibitor-Negative control (inhibitor-NC), miR-490-3p inhibitor, mimic- Negative control (mimic-NC) and miR-490-3p mimic group, and the transfection efficiency was verified by RT-qPCR. After 48 h of transfection, CCK8 assay, wound healing assay, Transwell invasion assay andflow cytometric were used to detect the effects of malignant biological characteristics such as proliferation, migration, invasion and apoptosis of SW1990 cell linerespectively. Western blot was used to detect the expression of E-cadherin and N-cadherin. Meanwhile, database predicted the target genes of miR-490-3p, and dual luciferase assay confirmed the targeted relationship between miR-490-3p and HMGA2, western blot was used to detect the expression of HMGA2. Results 1) The expression of miR-490-3p was downregulated in SW1990 cells (P = 0.215). 2) Overexpression of miR-490-3p significantly inhibited the proliferation (P < 0.0001), migration (P < 0.0001) and invasion (P < 0.0001) of SW1990 cell line and prompted apoptosis (P < 0.0001); In contrast, downregulation of miR-490-3p significantly promoted the proliferation (P < 0.0001), migration (P < 0.0001) and invasion (P < 0.0001) of SW1990 cell line and inhibited apoptosis (P < 0.0001). 3) Upregulation of miR-490-3p significantly decreased the expression of N-cadherin (P < 0.0001) and increased E-cadherin (P < 0.0001) expression in SW1990 cell line. In contrast, downregulation of miR-490-3p significantly upregulated (P < 0.0001) the expression of N-cadherin, decreased E-cadherin (P < 0.0001) expression. 4) HMGA2 was a target gene of miR-490-3p. Conclusion These findings indicate that miR-490-3p acts as a tumor suppressor during the process of EMT through targeting HMGA2, suggesting miR-490-3p is a potential new diagnostic and therapeutic target for the treatment of pancreatic cancer. [ABSTRACT FROM AUTHOR]

Objective: To investigate the effect of lentivirus-mediated microrna-376b-3p on the proliferation and apoptosis of pituitary adenomas and its possible mechanism. Methods: Taking human pituitary adenoma cell lines cultured in vitro as the research object, the microRNA-376b-3p mimics group (control group) and lenti-sh MicroRNA-376b-3p group (drug group) were established respectively, and the MTT method was used to detect MicroRNA-376b-3p on the proliferation of pituitary adenoma cells, the AnnexinV-FITC/PI double staining method was used to detect the effect of MicroRNA-376b-3p on the apoptosis rate of pituitary adenoma cells, and the Western blot method was used to detect the effect of MicroRNA-376b-3p on HMGA2, Bax, Caspase-3 protein expression. Result: (1) Lentivirus-mediated Microrna-376b-3p could significantly inhibit the proliferation of pituitary adenoma cells (P<0.05). (2) At 12, 24 and 48 h after microrna-376b-3p intervention, the apoptosis rate of pituitary gland cells in the control group was significantly lower than that in the drug group (P<0.05). Conclusions: (3) After transfection with microrna-376b-3p transformation, the expression of Bax protein increased, and the expression of Bcl-2, Caspase-3, Survivin and HMGA2 protein decreased (P<0.05). Lentivirus-mediated Microrna-376b-3p could significantly inhibit the proliferation of pituitary adenoma cells and induce apoptosis. The mechanism of microrna-376b-3p may be related to its targeted down-regulation of HMGA2 expression, up-regulation of Bax protein expression and regulation of survivin protein expression. [ABSTRACT FROM AUTHOR]

Objective: HMGA2(high-mobility group AT-hook 2) is a chromosome protein which has been well-documented to regulate cancer progression. We investigated whether and how HMGA2 affects Wnt signaling transduction process and the proliferation of colorectal cancer cells. Methods: RT-q PCR and western blot were utilized to examine m RNA and protein expression levels changes of HMGA2 in colorectal cancer tissues compared to nearby normal tissues. TOPflash reporter assay was conducted to detect the regulatory function of HMGA2 in Wnt signaling. Cell proliferation was quantified by Cell Counting Kit-8 in SW480 cells infected by adenovirus containing HMGA2 expression construct. Results: We found that both the m RNA and protein of HMGA2 were highly expressed in most of the examined human colorectal cancer samples. HMGA2 increased the expression of TOPflash a dose-dependent manner. And knockdown HMGA2 expression reduced Wnt3a-,Li Cl-,βcatenin- or Dvl(Dishevelled)-activated TOPflash-luciferase expression in colorectal cancer cell line HCT-116. Furthermore,ectopic expression of HMGA2 promoted proliferation of colorectal cancer cell line SW480. Conclusions: HMGA2 gene was highly expressed in the majority of the examined colorectal cancer samples. HMGA2 can promote the proliferation of colorectal cancer cells by up-regulating Wnt signaling. These results reveal the molecular and pathological functions of HMGA2 in colorectal cancer development and a potentially new regulatory effect on Wnt signaling. [ABSTRACT FROM AUTHOR]

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Background and objective HMGA2 is a small non-histone chromosomal protein. Recently, HMGA2 expression has been found in various malignant tumors and often correlates with the aggressiveness of the tumors. We investigated the expression of HMGA2 in the patients with non-small cell lung cancer and its clinical prognostic significance. Methods The expression level of HMGA2 in 59 patients with non-small cell lung cancer and 10 patients with benign lesions was detected by immunohistochemistry. Expression differences of HMGA2 and its relationship with the patients survival were analyzed with c2 test, Kaplan-Meier survival curve and Cox regression. Results There were 47 cases expressing HMGA2 (78％), positive rates for HMGA2 expression in pulmonary squamous, adenocarcinoma and squamous-adenocarcinoma were 70.4%, 82.1% and 100%, respectively. But there was no HMGA2 expression in the benign lesions. Differences of HMGA2 expression were significant between the two groups (P=0.013). It was also more highly expressed in the cases with lymphonode metastasis (P=0.007). Cox multivariate regression analysis showed that the disease stage was independently prognostic factors in NSCLC patients. Conclusion HMGA2 is overexpressed in the non-small cell lung cancer and related with disease stage and lymphonode metastasis. HMGA2 expression may be of prognostic importance in NSCLC.

Objective: To investigate whether miR-let-7c-5p inhibits invasion and migration of bladder cancer cells by regulating HMGA2., Methods: We used bioinformatics methods to determine the key genes of miR-let-7c-5p. RT-qPCR and Western blotting were used to detect the expressions of miR-let-7c-5p mRNA and HMGA2 protein in bladder cancer and adjacent tissues. With human normal bladder SV-HUC-1 cells as the control, we detected the expression levels of miR-let-7c-5p mRNA and HMGA2 protein in bladder cancer cell lines T24, UM-UC-3 and 5637 with RT-qPCR and Western blotting. We observed the effects of miR-let-7c-5p upregulation (by transfection with a miR-let-7c-5p mimic), miR-let-7c-5p downregulation (using a miR-let-7c-5p inhibitor), and knockdown of both HMGA2 and miR-let-7c-5p on invasion, migration and expressions of epithelial-mesenchymal transition (EMT)-related proteins (E-cadherin, N-cadherin, vimentin, and Snail) in UM-UC-3 cells. Dual luciferase assay, RT-qPCR and Western blotting were used to verify the targeting relationship between miR-let-7c-5p and HMGA2., Results: HMGA2 was identified as one of the target genes of miR-let-7c-5p. Compared with the adjacent tissues, bladder cancer tissues showed a significantly decreased expression of miR-let-7c-5p and an increased expression of HMGA2 protein ( P < 0.05). In UM-UC-3 cells, the expression of miR-let-7c-5p was significantly reduced and that of HMGA2 was significantly increased as compared with those in SV-HUC-1 cells ( P =0.001). Up-regulating miR-let-7c-5p expression significantly lowered the invasion and migration abilities of UM-UC-3 cells, and down-regulating miR-let-7c-5p expression obviously promoted the invasion and migration of UM-UC-3 cells ( P < 0.05). Knockdown of both miR-let-7c-5p and HMGA2 expression significantly lowered the invasion and migration ( P < 0.05) and inhibited the expressions of EMT-related proteins of UM-UC-3 cells ( P < 0.05)., Conclusion: miR-let-7c-5p inhibits EMT of bladder cancer UM-UC-3 cells by targeting HMGA2, thereby inhibiting the cell invasion and migration.