微小RNA活性分析法筛选前列腺癌去势抵抗转化相关微小RNA的实验研究

目的 探讨微小RNA(miRNA)活性分析法筛选前列腺癌去势抵抗转化相关miRNA的效率。方法 应用miRNA活性分析法筛选出在前列腺癌去势抵抗过程中潜在的发挥活性作用的miRNAs。培养人激素敏感型前列腺癌LNCAP细胞(对照组)及人去势抵抗型前列腺癌C4-2细胞(C4-2组)、PC-3细胞(PC-3组)、DU-145细胞(DU-145组),提取各组细胞总RNA,采用实时荧光定量PCR(qPCR)检测miRNAs,比较各组细胞miRNAs的表达情况。结果 应用miRNA活性分析法,依据筛选流程,共选出9个差异表达的miRNAs,分别为miR-1、miR-122、miR-218、miR-145、miR-155、miR-210、miR-197、 miR-346、let-7b。采用qPCR检测这9个miRNAs,结果显示,7个miRNAs在两种不同状态下的前列腺癌细胞中存在差异表达;在不同去势抵抗型前列腺癌细胞中,miR-210、miR-197、miR-346、miR-218均明显高表达,而miR-122、miR-145、let-7b均明显低表达。结论 采用miRNA活性分析法筛选前列腺癌去势抵抗转化相关miRNA,有着较高的准确性与可信度;其具体转化过程还需进一步证实。

Screening key micro RNAs for castration-resistant prostate cancer based on micro RNA activity analysis method

Objective To explore the efficiency of micro RNA(miRNA) activity analysis method in screening the miRNA associated with castration resistance in prostate cancer.Methods miRNAs were screened for their potential role in castration resistance of prostate cancer by miRNAs activity analysis method. Human hormone-sensitive prostate cancer LNCAP cells (control group) and castration-resistant prostate cancer C4-2 cells (C4-2 group), PC-3 cells (PC-3 group) and DU-145 cells (DU-145 group) were cultured. Total RNA was extracted from each group. Real-time fluorescence quantitative PCR (qPCR) was used to detect miRNAs and compare the expression of miRNAs in each group.Results According to the screening process, 9 differentially expressed miRNAs were identified by miRNA activity analysis method. They were miR-1, miR-122, miR-218, miR-145, miR-155, miR-210, miR-197, miR-346 and let-7b. The 9 miRNAs were detected by qPCR. The results showed that 7 miRNAs were differentially expressed in prostate cancer cells in two different states. In different castration-resistant prostate cancer cells, miR-210, miR-197, miR-346 and miR-218 were significantly over expressed, while miR-122, miR-145 and let-7b were significantly under expressed.Conclusions The screening of castration-resistant transformation-related miRNAs in prostate cancer by miRNAs activity analysis method has high accuracy and reliability, and the specific transformation process needs further confirmation.