NOV基因与Ki-67、VEGF、p27和COX-2在肾透明细胞癌中的相关性

NOV modulates expression of Ki-67,VEGF,p27 and COX-2 in renal clear cell carcinoma

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 research the correlation between nephroblastoma overexpressed gene（ NOV/CCN3 ） and vas- cular endothelial growth factor（ VEGF）, cyclin-dependent kinase inhibitor p27, cell proliferating antigen Ki-67, cycloox- ygenase-2 （COX-2）expression in renal clear cell carcinoma （ccRCC）. Methods The NOV expressed plasmid （pEG- FP-C1-NOV） was constructed and transfected into ccRCC cell line 786-0. The mRNA levels of VEGF, p27, Ki-67, COX-2, and NOV were tested via RT-PCR in 786-0 group, 786-O-empty vector group and 786-O-NOV group. Immu- nohistochemistry of NOV, Ki-67, VEGF, p27and COX-2 was performed on tissue microarrays （TMA） and immuno- histochemistry staining intensities were analyzed. Results NOV mRNA expression of 786-O-NOV group was higher compared with the other two groups, while there was no significant difference between another two groups. The 786-0- NOV cells expressed more p27, COX-2 and less Ki67, VEGF compared with the other two groups（ P 〈 0.05 ）. By im- munohistochemistry the expression of NOV was negatively related to the expressions of Ki67 and VEGF（ the correlation coefficients were - 0. 686 and - 0. 646, respectively, P 〈 0.05 ）, while positively to the expressions of p27 and COX- 2 （ the correlation coefficients were 0.491 and 0. 762 ,respectively, P 〈 0.05 ）. Conclusion NOV could influence the proliferation and invasion of tumor cells in ccRCC. The mechanism may be associated with its influence on the expressions of Ki-67, VEGF, p27 and COX-2.