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目的研究肾母细胞瘤过表达基因(NOV/CCN3)在肾透明细胞癌(ccRCC)中与增殖细胞核抗原Ki-67、血管内皮生长因子(VEGF)、细胞周期依赖性激酶抑制因子p27、环氧合酶2(COX-2)的相关性。方法用过表达NOV质粒pEGFP-C1-NOV转染人肾透明细胞癌细胞株786-O,实时定量PCR(RT-PCR)技术测定转染了过表达NOV质粒的细胞(实验组)、转染了空载体的细胞(空载组)及未行质粒转染的786-O细胞(空白组)中Ki-67、p27、VEGF和COX-2 mRNA表达的差异。应用免疫组化染色法对组织芯片行NOV及Ki-67、VEGF、p27、COX-2染色,得到量化资料并行相关性分析。结果 RT-PCR结果显示,实验组细胞与空白组细胞比较NOV mRNA水平增高(P<0.05),同时Ki-67、VEGF在mRNA水平降低(P<0.05),p27、COX-2 mRNA水平显著升高(P<0.05)。空白组与空载组相比较均无明显差异。免疫组化染色结果显示,肾透明细胞癌组织中NOV的表达与Ki-67、VEGF的表达呈负相关(r分别为-0.686、-0.646,P<0.05),而与p27、COX-2的表达呈正相关(r分别为0.491、0.762,P<0.05)。结论 NOV在肾透明细胞癌中对增殖和进展的作用可能与影响Ki-67、VEGF、p27、COX-2的表达相关。
Objective To investigate the relationship between the expression of NOV / CCN3 and the expression of proliferating cell nuclear antigen Ki-67, vascular endothelial growth factor (VEGF), cell cycle-dependent kinase inhibitor p27, Coenzyme 2 (COX-2) correlation. Methods Human renal clear cell carcinoma cell line 786-O was transfected with overexpression NOV plasmid pEGFP-C1-NOV. The cells transfected with NOV plasmid were detected by real-time quantitative PCR (RT-PCR) The differences of Ki-67, p27, VEGF and COX-2 mRNA expression in empty vector (empty group) and 786-O cells transfected without plasmid (blank group) The expression of NOV, Ki-67, VEGF, p27 and COX-2 in tissue microarray were detected by immunohistochemical staining, and the quantitative data were analyzed in parallel. Results The results of RT-PCR showed that the level of NOV mRNA in experimental group was significantly higher than that in blank group (P <0.05), while the expression of Ki-67 and VEGF was lower (P <0.05) and the level of p27 and COX-2 mRNA significantly increased High (P <0.05). No significant difference between blank group and no-load group. The results of immunohistochemistry showed that the expression of NOV in renal clear cell carcinoma was negatively correlated with the expression of Ki-67 and VEGF (r = -0.686, -0.646, P <0.05, respectively), but not with the expression of p27, COX- (R = 0.491,0.762, P <0.05). Conclusion The effect of NOV on the proliferation and progression of renal clear cell carcinoma may be related to the expression of Ki-67, VEGF, p27 and COX-2.