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To study the expression of cell cycle related factor Gli2 in autogenous vein graft and its relation with neointima formation.Autogenous vein graft model were established in 36 male wistar rats of 8 weeks old, 140 g, by transplanting the left jugular vein to intra renal abdominal aorta with microsurgical technique. Graft veins were harvest at 14, 28 days after transplantation. The IF and W-B were used to detect the protein expression in the vein graft. At the same time Gli2- mRNA was measured by RT-PCR.Immunofluorescent staining showed that the Gli2+ cells was only 3.2% ± 0.4% in the normal vein, but was much more in the vein graft after surgery, was 41.3% ± 0.6%, 58.3 ± 0.6% respectively; The expression of Gli2 and PCNA were both elevated in the vein graft. There is a positive correlation between them which indicated by W-B, the relation index was 0.826; the Gli2 mRNA content was also increased in vein graft, was 8.9, 13.6 fold compared with normal vein as 1 respectively.Gli2 is upregulated in autogenous vein grafts and may correlated with the proliferation of vascular smooth muscle cells.

To study the expression of cell cycle related factor sonic hedgehog (SHH) in autogenous vein graft and its relation with neointima formation.Autogenous vein graft model were established in 24 male Wistar rats of 8 weeks old and 140 g weight, by transplanting the left jugular vein to intra renal abdominal aorta with microsurgical technique. Graft veins were harvested at 14 d and 28 d after transplantation. The immunohistochemistry and Western blot were used to detect the SHH and PCNA expression in the vein graft. At the same time SHH mRNA was measured by quantitative real-time PCR. The opposite normal veins served as control.Histological staining showed that the percent of SHH+ cells was only (2.0 +/- 0.5)% in the normal vein, but was much more in the vein graft after surgery, as (39.4 +/- 0.4)% and (63.0 +/- 0.3)% respectively (P0.01). The expression of SHH and PCNA were both elevated in the vein graft. There was a positive correlation between them which indicated by Western blot (r = 0.808, P0.01). The SHH mRNA content also increased in vein graft to 9.5 and 23.8 folds of that in control.SHH is upregulated in autogenous vein grafts and may correlated with the proliferation of vascular smooth muscle cells.

To study the effect of shh on the migration, proliferation and phenotypic modulation of vascular adventitial fibroblasts.Cultivate the vascular adventitial fibroblast in vitro. Use immunofluorescent, laser confocal microscopy, Western-blot and real-time PCR to detect the expression of mRNA and protein of related index. Estimate cell proliferation according to the expression of Ki67 and cell proliferation curve. Application of wound healing test to estimate migration of fibroblast. The expression of alpha-actin is thought to be marker of phenotypic modulation of fibroblast.The expression of shh was detected in vascular adventitial fibroblast in vitro. After addition of exogenous shh (3.5 microg/ml), there were more Ki67(+) cells and the wounding area which was covered by cells became larger. The expression of alpha-actin was detected. After addition of cyclopamine (40 micromol/L), there were less Ki67(+) cells and the wounding area which was covered by cells became smaller.Shh can promote proliferation, migration and phenotypic modulation of vascular adventitial fibroblasts.

To assess the relationship between the XRCC1 polymorphism and susceptibility to lung cancer in non-smoking female on the basis of a hospital-based case-control study.Genotypes were determined by PCR-restriction fragment length polymorphism in 50 patients with lung cancer and 50 controls. The adjusted odds ratios (OR) and 95% confidence intervals (CI) were calculated using logistic regression model to study the relationship between different genotypes and risk of lung cancer in non-smoking women. Furthermore, a multiplicative interaction between exposure to cooking oil smoke and the variant XRCC1 399Gln allele on risk of lung adenocarcinoma was evaluated.Individuals carrying Gln/Gln genotype were at an increased risk to suffer from lung adenocarcinoma as compared with those with the Arg/Arg genotype (OR: 14.12; 95% CI: 2.14 approximately 92.95, adjusted for age and cooking oil smoke). The OR of lung adenocarcinoma for the variant XRCC1 399Gln allele with exposure to cooking oil smoke was 6.29 (95% CI 1.99 approximately 19.85).The above described findings indicate that Arg 399Gln polymorphism in the XRCC1 is associated with risk of lung adenocarcinoma but not with risk of squamous-cell carcinoma of the lung in non-smoking women.

T cell factor-4 (TCF-4) plays an important role in development and carcinogenesis. Recently, the role of TCF-4 has been described in colon cancer and other cancers. However, whether TCF-4 plays a similar role in lung cancer is unknown. To answer this question, we studied the expression of TCF-4 protein and mRNA in non-small-cell lung cancer (NSCLC) and the relation of TCF-4 expression pattern to histological type and cell differentiation.Tissue samples from sixty cases of pathologically diagnosed NSCLC and eight normal tissue samples were obtained between September 2001 and March 2003. Immunohistochemistry was used to investigate the distribution of TCF-4 protein. The staining patterns of the tumors were divided into 4 categories: nuclear staining alone or nuclear staining greater than cytoplasmic staining; cytoplasmic staining or cytoplasmic staining greater than nuclear staining; equal nuclear and cytoplasmic staining; no nuclear staining or cytoplasmic staining. The integrated optical density (OD) values of all sections were analyzed by UIC MetaMorph image analysis software. The expression of TCF-4 mRNA was detected by one-step reverse transcription-polymerase chain reaction (RT-PCR). The integrated density values of the PCR products were analyzed semi-quantitatively.Immunohistochemistry showed that there was no expression of TCF-4 in normal tissue. However, TCF-4 was expressed in 86.7% (52/60) of NSCLC samples, mainly in the nuclei of tumor cells. Furthermore, there was a significant difference in TCF-4 localization patterns between squamous cell carcinomas and adenocarcinomas (P0.05). The integrated OD values of TCF-4 expression was significantly higher in tumors with moderate-poor cell differentiation than in well differentiated tumors (51.63 +/- 6.67 vs 46.13 +/- 12.31, P0.01). There was no TCF-4 mRNA expression in normal tissue. However, 63.9% (23/36) of carcinoma samples expressed TCF-4 mRNA. TCF-4 mRNA expression was significantly higher in tumors with moderate-poor cell differentiation than in well differentiated tumors (P0.05). There were no significant differences in mRNA expression in comparison with histological type.The sub-cellular distribution of TCF-4 may correlate with NSCLC histological type. High expression of TCF-4 mRNA and protein may be associated with the degree of cell differentiation in NSCLC.

AIM: To study the effect of cyclin G2 on proliferation of gastric adenocarcinoma cell line-SGC-7901 cell in vitro. METHODS: By use of cation lipofectamine transfection reagent, the pIRES-G2 and pIRESneo plasmids were transferred into SGC-7901cell line. Anticlones were selected by G418. Positive clones were observed and counted using Giemsa staining. Cell proliferative ability was assayed by MTT. RESULTS: (1) The clone number of pIRES-G2 group decreased, clone volume reduced. The number of cell clones in pIRESneo group was 87 ± 3, that of pIRES-G2 group was 53 ± 4, occupying 60.1% of pIRESneo group, there was significant difference obviously (P < 0.01, t = 15.45). (2) The average absorbance of clone cell obtained by stable transfection of pIRES-G2 at 570 nm was 1.6966 ± 0.2125, the average absorbance of clone cell obtained by stable transfection of pIRESneo at 570 nm was 2.1182 ± 0.3675, there was significant difference between them (P < 0.01, t = 3.412). CONCLUSION: Cyclin G2 can inhibit SGC-7901cell proliferative ability obviously, it may be a negative regulator in cell cycle regulation.

Background T cell factor 4 (TCF 4) plays an importa nt role in development and carcinogenesis Recently, the role of TCF 4 has be en described in colon cancer and other cancers However, whether TCF 4 plays a similar role in lung cancer is unknown To answer this question, we studied the expression of TCF 4 protein and mRNA in nonsmallcell lung cancer (NSCLC) and the relation of TCF 4 express i on pattern to histological type and cell differentiation Methods Tissue samples f r om sixty cases of pathologically diagnosed NSCLC and eigh t normal tissue samples were obtained between September 2001 and March 2003 Immu n ohistochemistry was used to investigate the distribution of TCF 4 protein Th e s taining patterns of the tumors were divided into 4 categories: nuclear staining alone or nuclear staining greater than cytoplasmic staining; cytoplasmic stainin g or cytoplasmic staining greater than nuclear staining; equal nuclear and cytop lasmic staining; no nuclear staining or cytoplasmic staining The integrated op t ical density (OD) values of all sections were analyzed by UIC MetaMorph image an alysis software The expression of TCF 4 mRNA was detected by onestep rever s e t ranscriptionpolymerase chain reaction (RTPCR) The integrated density value s o f the PCR products were analyzed semiquantitatively Results Immunohistochemist ry showed that there was no expression of TCF 4 in normal tissue However, TC F  4 was expressed in 86.7% (52/60) of NSCLC samples, mainly in the nuclei of tumor cells Fu r thermore, there was a significant difference in TCF 4 localization patterns be t w een squamous cell carcinomas and adenocarcinomas (P005) The integrated OD val ues of TCF 4 expression was significantly higher in tumors with moderatepoor ce ll differentiation than in well differentiated tumors (5163±667 vs 4613 ±12 31, P001) There was no TCF 4 mRNA expression in normal tissue How e ver, 63.9% (23/36) of carcinoma samples expressed TCF 4 mRNA TCF 4 mRNA expression was signif ic ant ly higher in tumors with moderatepoor cell differentiation than in well differ e ntiated tumors (P005) There were no significant dif ferences in mRNA expression in comparison with histological type Conclusions The subcellular distribution of TCF 4 may correlate with NSCLC histological type High expres s io n of TCF 4 mRNA and protein may be associated with the degree of cell differen t iation in NSCLC