摘要
目的:探讨沉默高迁移率族蛋白B1(HMGB1)联合多西他赛(DTX)对前列腺癌细胞增殖和凋亡的影响及可能作用机制。方法:RT-qPCR法检测不同前列腺癌细胞系及正常前列腺上皮细胞HMGB1 mRNA表达;使用不同HMGB1小干扰RNA(si-HMGB1-1、si-HMGB1-2、si-HMGB1-3)转染PC-3细胞,检测沉默效果;MTT法检测不同浓度DTX对PC-3细胞增殖的影响;PC-3细胞随机分为对照组(常规培养)、si-HMGB1-NC组(转染si-HMGB1-NC)、si-HMGB1组(转染si-HMGB1-3)、DTX组(20 nmol/L DTX)、si-HMGB1+DTX组(转染si-HMGB1-3+20 nmol/L DTX),MTT法检测细胞存活率;流式细胞术检测细胞凋亡率;Western印迹法检测细胞中HMGB1、B细胞淋巴瘤-2(Bcl-2)、Bcl-2相关X蛋白(Bax)蛋白表达。结果:PC-3细胞中HMGB1 mRNA表达最高;转染si-HMGB1-3后HMGB1 mRNA表达最低;各浓度DTX均可抑制PC-3细胞增殖。与对照组比较,si-HMGB1组、DTX组A值、细胞存活率、HMGB1、Bcl-2蛋白表达下降,凋亡率、Bax蛋白表达升高(P<0.05);与si-HMGB1组、DTX组比较,si-HMGB1+DTX组A值、细胞存活率、Bcl-2蛋白表达下降,凋亡率、Bax蛋白表达升高,且si-HMGB1+DTX组HMGB1蛋白表达较DTX组降低(P<0.05)。结论:沉默HMGB1联合DTX化疗可抑制前列腺癌细胞增殖,促进细胞凋亡,可能与调控Bcl-2家族相关蛋白表达有关。
Objective: To investigate the effect of silencing the high-mobility group box-1 protein(HMGB1) combined with docetaxel(DTX) on the proliferation and apoptosis of PCa cells and its possible action mechanism. Methods: The expression of HMGB1 mRNA in different PCa cell lines and normal prostatic epithelial cells was detected by RT-qPCR. The PC-3 cells were transfected with different HMGB1 small interfering RNAs(si-HMGB1, si-HMGB1-2 and si-HMGB1-3), and the silencing effect was detected. The effects of different concentrations of DTX on the proliferation of the PC-3 cells was determined by MTT. Then the PC-3 cells were randomly divided into five groups: control(conventional culture), si-HMGB1-NC(si-HMGB1-NC transfection), si-HMGB1(si-HMGB1-3 transfection), DTX(20 nmol/L DTX), and si-HMGB1+DTX(si-HMGB1-3+20 nmol/L DTX transfection), followed by measurement of the survival rate of the cells by MTT, their apoptosis rate by flow cytometry, and the expressions of HMGB1, B-cell lymphoma-2(Bcl-2) and Bcl-2-associated X(Bax) proteins in different groups by Western blot. Results: The expression of HMGB1 mRNA in the PC-3 cells was the highest and the lowest after transfection with si-HMGB1-3. DTX inhibited the proliferation of the PC-3 cells at various concentrations. Compared with the control group, the si-HMGB1 and DTX groups showed significantly decreased A values, cell survival rates and HMGB1 and Bcl-2 expressions, but increased cell apoptosis rates and Bax expressions(P < 0.05). In comparison with the si-HMGB1 and DTX groups, the si-HMGB1+DTX group exhibited a remarkably decreased A value, cell survival rate and Bcl-2 expression, but increased cell apoptosis and Bax expression. The expression of the HMGB1 protein was markedly lower in the si-HMGB1+DTX than in the DTX group(P < 0.05). Conclusion: Silencing HMGB1 combined with DTX chemotherapy can inhibit the proliferation and promote the apoptosis of PCa cells, which may be attributed to its regulatory effect on the expressions of the Bcl-2 family-related proteins.
作者
冯武兵
陈晨
FENG Wu-bing;CHEN Chen(Department of Urology,Central Hospital of Chongqing University,Chongqing 400016,China;Shangqingsi Street Community Health Service Center of Yuzhong District,Chongqing 400015,China)
出处
《中华男科学杂志》
CAS
CSCD
北大核心
2021年第10期876-881,共6页
National Journal of Andrology
关键词
前列腺癌
高迁移率族蛋白B1
多西他赛
增殖
凋亡
prostate cancer
high-mobility group protein B1
docetaxel
proliferation
apoptosis
