miRNA-592在肾细胞癌进展中作用及其与预后关系研究

The role of miRNA-592 in the progression of renal cell carcinoma and its relationship with prognosis

目的探讨miRNA-592在肾细胞癌(RCC)中的作用及其与预后的关系。方法逆转录-聚合酶链反应检测110例RCC组织及其配对的癌旁组织、正常人肾上皮细胞系(HK-2)和人RCC细胞系(786-O、ACHN、Caki-1和769-P)中miRNA-592的表达。分析miRNA-592与RCC临床病理特征及生存预后的关系,生存分析用Kaplan-Meier法。用LipofectAMINETM 2000试剂盒将miRNA-592沉默质粒(sh-miRNA-592组)或空载体质粒(sh-NC组)转染入786-O细胞,克隆形成实验和CCK-8法检测增殖能力,Annexin-V-FITC/PI流式细胞术检测细胞凋亡,蛋白免疫印迹法检测细胞增殖相关蛋白(Cyclin B1、增殖细胞核抗原)和凋亡相关蛋白(Bax、Cleaved caspase-3)。结果癌组织中miRNA-592相对表达量为(2.34±0.47),明显高于癌旁组织的(0.97±0.41),HK-2细胞的miRNA-592相对表达量为(1.01±0.08),低于786-O细胞(3.84±0.47)、ACHN细胞(2.76±0.53)、Caki-1细胞(1.98±0.12)与769-P细胞(2.45±0.52),差异均有统计学意义(P<0.05)。miRNA-592高、低表达组的T分期患者比例比较,差异有统计学意义(P<0.05)。miRNA-592高表达患者的存活率低于低表达患者,差异有统计学意义(P<0.05)。sh-miRNA-592组miRNA-592相对表达量、细胞克隆数与细胞培养48、72和96 h时细胞增殖检测光密度值均低于sh-NC组,sh-miRNA-592组细胞凋亡率高于sh-NC组,差异均有统计学意义(P<0.05)。结论miRNA-592在RCC中的高表达与不良生存预后有关,沉默miRNA-592可抑制RCC细胞的增殖并促进凋亡。

Objective To investigate the role of miRNA-592 in renal cell carcinoma(RCC)and its relationship with prognosis.Methods Reverse transcription-polymerase chain reaction detection was performed on miRNA-in 110 cases of RCC tissues and their paired adjacent tissues,normal human renal epithelial cell lines(HK-2)and human RCC cell lines(786-O,ACHN,Caki-1 and 769-P)miRNA-592 expression.To analyze the relationship between miRNA-592 and RCC clinicopathological characteristics and survival prognosis,the Kaplan-Meier method was used for survival analysis.LipofectAMINETM 2000 kit was used to transfect miRNA-592 silencing plasmid(sh-miRNA-592 group)or empty vector plasmid(sh-NC group)into transfected 786-O cells,clone formation experiment and CCK-8 method were used to detect proliferation ability,Annexin-V-FITC/PI flow cytometry was used to detect cell apoptosis,Western-blot was used to detect cell proliferation-related proteins(Cyclin B1,proliferating cell nuclear antigen)and apoptosis-related proteins(Bax,Cleaved caspase-3).Results The relative expression level of miRNA-592 in cancer tissues was(2.34±0.47),which was significantly higher than that in adjacent tissues(0.97±0.41).The relative expression level of miRNA-592 in HK-2 cells was(1.01±0.08),which was lower than that of 786-O cells(3.84±0.47),ACHN cells(2.76±0.53),Caki-1 cells(1.98±0.12)and 769-P cells(2.45±0.52),and the differences were statistically significant(P<0.05).There were statistically significant differences in the proportion of T stage patients in the high and low miRNA-592 expression groups(P<0.05).The overall survival rate of patients with high miRNA-592 expression was lower than that of patients with low miRNA-592 expression(P<0.05).The relative expression of miRNA-592,the number of cell clones and the optical density of cell proliferation detected at 48,72 and 96 hours in SH-NC group were lower than those in SH-NC group,and the apoptosis rate in SH-miRNA-592 group was higher than that in SH-NC group(P<0.05).Conclusion The high expression of miRNA-592 in RCC is related to poor survival prognosis,silencing miRNA-592 can inhibit the proliferation of RCC cells and promote apoptosis.