STAG2基因在肾癌的表达及功能研究

Expression and function of STAG2 gene in renal cell carcinoma

目的研究骨髓基质抗原2(STAG2)基因在人肾癌组织和肾癌细胞中的表达水平并研究STAG2基因对肾癌细胞生物学表型的影响。方法使用实时荧光定量PCR(qRT-PCR)检测STAG2基因在5株肾癌细胞、1株正常肾细胞和68对肾癌及其对应的癌旁组织中的表达水平;使用Western blot法分析STAG2蛋白在5株肾癌细胞、1株正常肾细胞的表达特点;使用慢病毒包装感染ACHN、786-O细胞并使用qRT-PCR检测感染效率;CCK8试剂盒检测STAG2过表达后对肾癌细胞增殖的影响;Transwell法检测STAG2过表达后对肾癌细胞侵袭的影响;流式细胞术检测STAG2过表达后对肾癌细胞凋亡的影响。结果 (1)qRT-PCR结果显示,与正常肾细胞相比,STAG2基因在5株肾癌细胞的表达显著下降,差异有统计学意义(P<0.01);(2)Western blot结果显示在5株肾癌细胞中STAG2蛋白表达明显低于正常肾细胞;(3)qRT-PCR检测肾癌及其癌旁组织的STAG2表达水平结果显示与其对应癌旁组织相比,72.06%(49/68)肾癌组织中STAG2表达水平下降(P<0.01);肾癌组织中STAG2表达水平与肿瘤的临床病理分期相关(P=0.029),与患者年龄、性别、肿瘤组织学分级无明显相关性;(4)慢病毒感染ACHN、786-O细胞后STAG2表达水平明显升高(P<0.01);(5)体外实验中,过表达STAG2基因明显抑制肾癌细胞的增殖和迁移能力,促进肾癌细胞的早期凋亡(P<0.05,P<0.01)。结论 (1)在肾癌细胞和肾癌组织中STAG2的表达水平均有明显的下降,提示STAG2可能参与肾癌的发生和发展的过程,STAG2低表达对肾癌的早期诊断可能提供一些帮助;(2)上调STAG2基因的表达明显抑制肾癌细胞的增殖和迁移能力、促进肾癌细胞的凋亡,提示STAG2可能成为治疗肾癌的一个潜在靶点。

Objective To investigate the expression of stromal antigen 2 （STAG2） in human renal carcinoma cells and normal renal cells, and explore its impact on the biology behaviors of renal carcinoma cells. Methods The relative expression level of STAG2 was measured in renal cancer and pair-matched adjacent normal renal tissues, STAG2 gene was detected by quantitative real-time reverse transcription PCR（qRT-PCR） in five renal carcinoma cells, one normal kidney cell, 68 pairs of renal carcinoma and pair-matched adjacent normal renal tissues, the experimental results were statistically analyzed; the expression of STAG2 in 5 renal cell carcinoma cell lines and normal renal cell line were analyzed by Western blot; ACHN and 786-0 were infected with lentivirus packaging and the efficiency of infection was detected by qRT-PCR ; The effect of STAG2 overexpression on the proliferation of renat carcinoma cells was detected by CCK8 kit; the effect of STAG2 overexpression on the migration of renal carcinoma was detected by Transwell ; Flow cytometry was used to detect the apoptosis of cancer cells. Results （1） The results of qRT-PCR showed that the expression of STAG2 gene in 5 renal carcinoma cell lines was significantly lower than that of normal kidney cell line （ P 〈 0. 01 ） ; （2） The results of Western blot showed that the expression of STAG2 in 5 renal carcinoma cell lines were significantly lower than that in normal renal cell line; （3） The qRT-PCR results showed that STAG2 expression was decreased in 72.06% （49/68） renal carcinoma tissues compared with that in adjacent tissues（P 〈0. 01 ） and the expression level of STAG2 in renal carcinoma was correlated with the clinical pathological stage （ P = 0. 029 ） ; there was no significant correlation between age, sex and tumor histological grade; （4） STAG2 expression in ACHN and 786-0 were significantly increased using lentivirus infection （P 〈0. 01 ） ; （5） STAG2 overexpression attenuated cell proliferation, migration and promoted apoptosis in renal cell lines （ P 〈 0. 05, P 〈 0. 01 ）. Conclusion （1） The expression level of STAG2 in renal carcinoma cells and renal carcinoma tissues are significantly decreased, suggesting that STAG2 may be involved in the development and progression of renal cell carcinoma. （2） Low expression of STAG2 may be helpful in the early diagnosis of renal cell carcinoma; STAG2 overexpression attenuated cell proliferation, migration and promoted apoptosis in renal cell lines, suggesting that STAG2 may be a potential target for the treatment of renal cell carcinoma.