miR-195靶向抑制S6K1基因对前列腺癌侵袭和转移的影响

发布时间：2018-03-22 06:17

  本文选题：前列腺癌　切入点：miR-　出处：《实用医学杂志》2017年22期 　论文类型：期刊论文

【摘要】：目的研究miR-195靶向抑制S6K1基因调控前列腺癌细胞增殖、凋亡、侵袭和转移活性。方法选择2种前列腺癌细胞株DU145和LNCaP,实验分为3组,即空白对照组、过表达组和低表达组,构建DU145和LNCaP过表达和沉默表达miR-195的细胞株,分别培养24 h,采用反转录PCR(RT-PCR)法鉴定转染效果,MTT法检测细胞增殖率,流式细胞术检测凋亡率,Transwell侵袭实验和细胞划痕实验检测细胞侵袭和转移能力;RT-PCR法检测S6K1 mRNA相对表达水平,荧光素酶报告系统确认miR-195直接调控的靶基因。结果过表达组细胞增殖率显著低于对照组,低表达组最高;凋亡率高于对照组,低表达组最低;侵袭细胞数目和划痕距离小于对照组,低表达组最大;S6K1 mRNA相对表达水平低于对照组,低表达组最高。S6K1基因为miR-195的靶基因。结论 miR-195靶向抑制S6K1基因调控前列腺癌细胞的增殖、凋亡、侵袭和转移活性。
[Abstract]:Objective to study the inhibitory effect of miR-195 on the proliferation, apoptosis, invasion and metastasis of prostate cancer cell line DU145 and LNCaP.Methods two kinds of prostate cancer cell lines DU145 and LNCaP were divided into three groups: blank control group, overexpression group and low expression group. DU145 and LNCaP overexpression and silencing expression of miR-195 were constructed and cultured for 24 h respectively. The transfection effect was evaluated by reverse transcription PCR RT-PCR assay. The apoptosis rate was detected by flow cytometry and the relative expression of S6K1 mRNA was detected by reverse transcription-polymerase chain reaction (RT-PCR). Luciferase reporting system confirmed the target gene directly regulated by miR-195. Results the cell proliferation rate of overexpression group was significantly lower than that of control group, the highest in low expression group, the apoptosis rate was higher than that in control group, and the lowest in low expression group. The relative expression level of S6K1 mRNA in the low expression group was lower than that in the control group, and the highest S6K1 gene in the low expression group was due to the target gene of miR-195. Conclusion miR-195 can inhibit the proliferation of prostate cancer cells regulated by S6K1 gene. Apoptosis, invasion and metastasis activity.
【作者单位】： 南华大学附属第二医院泌尿外科;南华大学附属南华医院麻醉科;
【基金】：湖南省衡阳市科技计划项目(编号:2016KJ32)
【分类号】：R737.25
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本文编号：1647428