沉默液泡型ATP酶c亚基ATP6V0C抑制人前列腺癌细胞侵袭的分子机制

发布时间：2018-06-29 04:17

本文选题：液泡型ATP酶 + ATPVC　；参考：《北京大学学报(医学版)》2017年06期

【摘要】：目的:采用RNA干涉技术沉默液泡型ATP酶c亚基ATP6V0C的表达,探索ATP6V0C在前列腺癌侵袭中的分子作用机制,并研究其与LASS2/TMSG1的关系。方法与结果:ATP6V0C在高转移潜能人前列腺癌细胞系(PC-3M-1E8和PC-3M)中的表达明显高于低转移潜能人前列腺癌细胞系(PC-3M-2B4和PC-3),选择ATP6V0C表达量最高的PC-3M-1E8细胞进行基因沉默实验,发现ATP6V0C的siRNA转染PC-3M-1E8后,其基质金属蛋白酶2(matrix metalloprotein-2,MMP-2)和基质金属蛋白酶9(MMP-9)蛋白的表达量及MMP-2的分泌量均无明显变化,但上清液中MMP-9的活性明显减弱,与空白对照组及阴性对照组相比,差异有统计学意义(P0.05);干扰组细胞外氢离子浓度及液泡型ATPase的活性明显降低(P0.05);细胞划痕修复实验及transwell体外侵袭实验结果显示ATP6V0C siRNA干扰组细胞的体外迁移及侵袭能力明显减弱(P0.05)。PC-3M-1E8细胞转染ATP6V0C siRNA后,LASS2/TMSG1的表达明显减弱(P0.05)。免疫荧光双重染色结果显示ATP6V0C蛋白与LASS2/TMSG1共定位于胞浆和胞膜,干扰组共定位信号与对照组相比明显减弱。结论:特异性siRNA沉默ATP6V0C能抑制人前列腺癌细胞的迁移和侵袭能力,其机制与ATP6V0C沉默后抑制V-ATPase的活性,使细胞外氢离子浓度降低,抑制MMP-9的激活,从而影响细胞外基质的降解和重塑有关。靶向siRNA干扰ATP6V0C的表达后,可能通过反馈调节机制抑制LASS2/TMSG1的表达,但其具体分子机制尚待进一步研究。
[Abstract]:Aim: to silence the expression of the vacuolar ATPase c subunit ATP-6V0C by RNA interference technique, and to explore the molecular mechanism of ATP-6V0C in the invasion of prostate cancer, and to investigate its relationship with LASS2 / TMS320G1. Methods and results the expression of ATP 6V0C in high metastatic potential human prostate cancer cell lines (PC-3M-1E8 and PC-3M) was significantly higher than that in low-metastatic human prostate cancer cell lines (PC-3M-2B4 and PC-3). It was found that the expression of matrix metalloprotein-2 (MMP-2) and matrix metalloproteinase-9 (MMP-9) and the secretion of MMP-2 were not significantly changed after transfection of ATP 6V0C siRNA into PC-3M-1E8, but the activity of MMP-9 in the supernatant was significantly decreased, compared with the control group and the negative control group. The difference was statistically significant (P0.05), the extracellular hydrogen ion concentration and the activity of vacuolar ATPase were significantly decreased in the interference group (P0.05). The results of cell scratch repair and transwell invasion in vitro showed that the extracellular migration and invasion ability of the cells in vitro were observed in the ATT6V0C siRNA interference group (P0.05). PC-3M-1E8 cells were transfected with ATP-6V0C siRNA and the expression of LASS2 / TMS320G1 was significantly decreased (P0.05). The results of immunofluorescence double staining showed that ATP 6V0C protein and LASS2 / MS G1 co-located in the cytoplasm and membrane, and the co-localization signal in the interference group was significantly lower than that in the control group. Conclusion: the specific siRNA silencing ATP-6V0C can inhibit the migration and invasion of human prostate cancer cells. The mechanism of inhibition of V-ATPase activity, the decrease of extracellular hydrogen ion concentration, and the activation of MMP-9 after the silencing of ATP 6V0C. Thus affecting the degradation and remodeling of extracellular matrix. After targeted siRNA interference, the expression of ATP 6V0C may be inhibited by feedback regulation mechanism, but its molecular mechanism needs to be further studied.
【作者单位】：北京大学基础医学院病理学系;青岛中心医院病理科;内蒙古医学院基础医学院病理学系;内蒙古医学院附属医院病理科;北京科技大学机械学院;首都师范大学附属育新学校;
【基金】：国家自然科学基金(81572533)资助~~
【分类号】：R737.25

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