沉默前列腺特异性膜抗原促进前列腺癌LNCAP细胞上皮-间质转化及迁移、侵袭的研究

doi:10.3969/j.issn.1009-976X.2023.01.010

岭南现代临床外科 ›› 2023, Vol. 23 ›› Issue (01): 58-63.DOI: 10.3969/j.issn.1009-976X.2023.01.010

沉默前列腺特异性膜抗原促进前列腺癌LNCAP细胞上皮-间质转化及迁移、侵袭的研究

李金^1,4, 赖义明^2,3,4, 曾乐祥^2,3

1.中山大学孙逸仙纪念医院生殖中心,广州 510120;
2.中山大学孙逸仙纪念医院泌尿外科,广州 510120;
3.广东省泌尿系统疾病临床医学研究中心,广州 510120;
4.广东省恶性肿瘤表观遗传与基因调控重点实验室,广州 510120

通讯作者: *曾乐祥,Email：zenglx3@mail.sysu.edu.cn
基金资助:
国家自然科学基金资助项目（81802527）; 广东省省级科技计划项目（2021A1515010223）; 白求恩泌尿肿瘤专项研究基金（mnzl202026）; 广东省恶性肿瘤表观遗传与基因调控重点实验室基金（2020B1212060018）; 广东省泌尿系统疾病临床医学研究中心（2020B1111170006）

Knockdown of PSMA expression promoted cell migration, invasion and EMT in prostate cancer LNCAP cells

LI Jin^1,4, LAI Yi-min^2,3,4, ZENG Le-xiang^2,3,4

1. Reproductive Center, Sun Yat Sen Memorial Hospital, Sun Yat Sen University, Guangzhou 510120, China;
2. Urology, Sun Yat Sen Memorial Hospital, Sun Yat Sen University, Guangzhou 510120, China;
3. Guangdong Clinical Medical Research Center for Urinary Diseases, Guangzhou 510120, China;
4. Guangdong Provincial Key Laboratory of Malignant Tumor Epigenetics and Gene Regulation, Guangzhou 510120, China

Received:2022-10-09 Online:2023-02-20 Published:2023-04-13
Contact: ZENG Le-xiang, zenglx3@mail.sysu.edu.cn

摘要/Abstract

摘要： 目的利用RNAi技术沉默LNCAP细胞中前列腺特异性膜抗原（PSMA）表达并检测表达情况,检测沉默PSMA后LNCAP细胞迁移及侵袭等生物学行为的变化情况,以及LNCAP细胞中上皮-间质转化标志蛋白的变化情况。方法培养LNCAP细胞,分为si-PSMA组,阴性对照（negative control siRNA）组,抑制剂LY294002组和LY294002+si-PSMA组,采用qRT-PCR 和Western blot技术检测沉默PSMA后LNCAP细胞中PSMA的mRNA和蛋白表达情况,通过Transwell小室穿透实验检测LNCAP细胞迁移及侵袭能力改变,通过Western blot蛋白印迹法检测E-cadherin、β-cadherin、vimentin,snail等细胞上皮-间质转化标志蛋白的表达情况以及p-Akt（ser473）蛋白表达情况。结果与阴性对照组相比,si-PSMA组PSMA的mRNA和蛋白表达水平都明显降低（P<0.05）,Transwell结果显示迁移及侵袭细胞增多（P<0.01）,LY294002组细胞迁移及侵袭下调（P<0.05）,而LY294002+si-RNA组则可逆转下降趋势（P<0.05）,Western blot结果显示E-cadherin与β-cadherin的表达减弱,而vimentin、snail和p-Akt（ser473）的表达增强。结论沉默PSMA可能通过激活上皮间质化通路促进前列腺癌细胞迁移及侵袭能力。

关键词: 前列腺特异性抗原蛋白, 前列腺癌, 上皮-间质转化

Abstract: Objective To investigate the PSMA expression, migration, invasion and expression of EMT associated proteins in LNCAP cells after transfection of PSMA siRNA. Methods LNCAP cells were cultured and divided into si-PSMA group, negative control siRNA group, inhibitor LY294002 group and LY294002+si-PSMA group. The expressions of PSMA mRNA and protein were detected respectively by qRT-PCR and Western blot. Transwell assay were used to detect migration and invasion in LNCAP cells.Western blot was used to detect the effect of PSMA inhibitionon the protein expression of p-Akt （ser473） and epithelial mesenchymal transformation marker proteins such as E-cadherin, β-cadherin, vimentin and snail. Results Compared with negative control group, the mRNA and protein expression of PSMAin LNCAP cells was downregulated by transfection of PSMAsiRNA（P<0.05）. Transwell results showed that the migration and invasion rates were increased in si-PSMA group（P<0.05） and decreased in LY294002 group,but was rescued combined with siRNA（P<0.05）. Western blot results displayed higher expression of vimentin, snail and p-Akt（ser473） while lower expression of E-cadherin and β-cadherin in si-PSMA group. Conclusion Knockdown of PSMA may promote the migration and invasion of LNCAP cells by activating epithelial mesenchymal transformation pathway.

Key words: PSMA, prostate cancer, EMT

中图分类号:

R737.25

李金, 赖义明, 曾乐祥. 沉默前列腺特异性膜抗原促进前列腺癌LNCAP细胞上皮-间质转化及迁移、侵袭的研究[J]. 岭南现代临床外科, 2023, 23(01): 58-63.

LI Jin, LAI Yi-min, ZENG Le-xiang. Knockdown of PSMA expression promoted cell migration, invasion and EMT in prostate cancer LNCAP cells[J]. Lingnan Modern Clinics In Surgery, 2023, 23(01): 58-63.

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沉默前列腺特异性膜抗原促进前列腺癌LNCAP细胞上皮-间质转化及迁移、侵袭的研究

Knockdown of PSMA expression promoted cell migration, invasion and EMT in prostate cancer LNCAP cells

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