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目的检测肾癌及癌旁配对组织、肾癌及正常肾细胞系中mi R-15a-5p的表达水平,比较配对组织及细胞系之间的表达水平,分析表达水平与患者临床特征之间的关系,探究mi R-15a-5p作为肾癌标记物应用于临床的可能性。方法提取肾癌及正常肾细胞系、36对肾癌与配对癌旁组织标本中的总RNA,经反转录、q PCR检测mi R-15a-5p的表达,使用统计软件分析组织间、细胞系之间mi R-15a-5p的表达差异,分析mi R-15a-5p表达水平与患者临床特征间的关系。通过转染mi R-15a-5p干扰敲低肾癌细胞中mi R-15a-5p表达水平,利用流式细胞术评估其对肾癌细胞的凋亡影响。利用SPSS 19.0进行配对t检验及Fisher确切概率检验。结果 mi R-15a-5p在肾癌组织中表达水平明显高于癌旁正常组织(P<0.05),癌组织中表达水平为癌旁组织的(9.95±0.92)倍。肾癌细胞系(786-O、ACHN)中mi R-15a-5p表达量分别是正常肾细胞系(HEK-293T)的(8.40±1.74)倍、(2.94±0.11)倍(P<0.05)。标本中共有26例(72.2%)肾癌标本mi R-15a-5p表达上调,癌组织中mi R-15a-5p表达水平与肾癌临床分期相关(P<0.05)。敲低mi R-15a-5p表达水平可诱导肾癌细胞凋亡,786-O细胞凋亡率为(14.01±0.81)%(敲低mi R-15a-5p)和(2.47±0.28)%(阴性对照)(P<0.05),ACHN细胞凋亡率为(20.30±0.47)%(敲低mi R-15a-5p)和(11.45±0.61)%(阴性对照)(P<0.05)。结论 mi R-15a-5p在肾癌组织及细胞系中表达明显上调,且和肾癌分期相关,mi R-15a-5p可调节肾癌细胞凋亡,提示其在肾癌的发生发展中起着一定的作用,有作为肾癌标记物应用于临床的可能。
Objective To detect the expression of mi R-15a-5p in renal cell carcinoma and paracancer tissues, renal cell carcinoma and normal renal cell lines, compare the expression levels of paired tissues and cell lines, and to analyze the relationship between expression level and clinical features of patients Relationship, to explore the possibility of mi R-15a-5p as a marker of kidney cancer for clinical application. Methods The total RNA was extracted from renal cell carcinoma and normal renal cell lines and from 36 pairs of adjacent tissues of renal cell carcinoma and matched paracancerous tissues. The expression of mi R-15a-5p was detected by reverse transcription and q-PCR. The relationship between the expression level of mi R-15a-5p and the clinical features of the patients was analyzed. The effect of mi R-15a-5p transfection on the expression of mi R-15a-5p in renal cell carcinoma was evaluated by flow cytometry. Paired t-test and Fisher’s exact test were performed using SPSS 19.0. Results The expression of mi R-15a-5p was significantly higher than that in adjacent normal tissues (P <0.05). The expression level of mi R-15a-5p was 9.95 ± 0.92 times of the adjacent normal tissues. The expression level of mi R-15a-5p in renal cancer cell line (786-O, ACHN) was (8.40 ± 1.74) times and (2.94 ± 0.11) times higher than that in normal renal cell line (HEK- . The expression of mi R-15a-5p was up-regulated in 26 (72.2%) renal cancer specimens. The expression of mi R-15a-5p in cancer tissues was correlated with the clinical stage of renal cell carcinoma (P <0.05). Overexpression of mi R-15a-5p knocked down the expression of mi R-15a-5p and induced the apoptosis of 786-O cells. The apoptotic rates of 786-O cells were (mi.01-15.1%) (P <0.05). The apoptosis rate of ACHN cells was (20.30 ± 0.47)% (knockdown mi R-15a-5p) and (11.45 ± 0.61)% (negative control) Conclusion The expression of mi R-15a-5p is significantly upregulated in renal cell carcinoma and cell lines, and is related to the stage of renal cell carcinoma. Mi R-15a-5p can regulate the apoptosis of renal cell carcinoma, suggesting that it plays an important role in the development of renal cell carcinoma With a certain role, as a marker of kidney cancer may be used in clinical.