Mesothelin对乳腺癌细胞MDA-MB-231生物学行为的影响

Effects of Mesothelin on proliferation,apoptosis,migration and invasion of breast cancer MDA-MB-231 cells

目的探讨间皮素(Mesothelin)对三阴性乳腺癌(TNBC)细胞增殖、凋亡、迁移及侵袭能力的影响。方法免疫化学染色检测TNBC组织、癌旁组织及乳腺癌细胞系中Mesothelin的表达。将过表达Mesothelin慢病毒质粒(Mesothelin组)及对照Scramble质粒(Scramble组)转染TNBC细胞系MDA-MB-231细胞,未感染细胞作为Control组。流式细胞术检测病毒感染效率;实时荧光定量PCR、流式细胞术及Western blot检测Mesothelin表达;CCK-8检测细胞增殖情况;利用Annexin V-APC/PI检测细胞凋亡;Western blot检测B淋巴细胞瘤-2(Bcl-2)、Bcl-2关联X蛋白(Bax)、E-钙黏蛋白(E-cadherin)、N-钙黏蛋白(N-cadherin)、波形蛋白(Vimentin)表达;划痕实验检测细胞迁移能力;Transwell实验检测细胞的侵袭能力。结果Mesothelin在TNBC肿瘤组织中表达高于癌旁组织(P<0.05),在乳腺癌细胞系中有表达。流式细胞术验证发现慢病毒感染效率达到90%以上。与Scramble组相比,Mesothelin组Mesothelin mRNA和蛋白表达水平升高,细胞增殖率升高,细胞凋亡率降低,Bax、E-cadherin蛋白表达减少,Bcl-2、Ncadherin和Vimentin蛋白表达增加,细胞的迁移及侵袭能力增强(P<0.05)。结论Mesothelin能够促进乳腺癌细胞增殖,抑制细胞凋亡,增加细胞的迁移及侵袭能力。

Objective To investigate the effect of Mesothelin on proliferation,apoptosis,migration and invasion of human triple-negative breast cancer cells(TNBC).Methods Immunohistochemical staining was used to detect the expression of Mesothelin in TNBC tissue,paracancer tissue and breast cancer cell lines.TNBC cell line MD-MB-231 cells were transfected with the overexpressed Mesothelin lentivirus plasmid(the mesothelin group)and the control Scramble plasmid(the Scramble group).MD-MB-231 cells with no infected lentivirus were used as the control group.Flow cytometry was used to detect the efficiency of virus infection.The expression of Mesothelin was detected by qPCR,flow cytometry and Western blot assay.Cell proliferation was detected by CCK-8.Annexin V-APC/PI was used to detect apoptosis.The expression of anti-B lymphoblastoma-2(Bcl-2),anti-Bcl-2 associated X protein(Bax),E-cadherin,N-cadherin and Vimentin were detected by Western blot assay.Cell migration ability was detected by scratch test.The invasive ability of cells was detected by Transwell assay.Results The expression of Mesothelin was significantly higher in TNBC tumor tissue than that in paracancer tissue(P<0.05),and Mesothelin was expressed in breast cancer cell line.Flow cytometry showed that the efficiency of lentivirus infection was more than 90%.Compared with the Scramble group,the expression level of Mesothelin mRNA was increased in the Mesothelin group,and the cell proliferation rate was increased,while the cell apoptosis rate was decreased.The expression of Bax and E-cadherin protein decreased,the expression of Bcl-2,N-cadherin and Vimentin protein increased,and the migration and invasion ability of cells were significantly enhanced(P<0.05).Conclusion Mesothelin can promote the proliferation,inhibit the apoptosis and increase the ability of migration and invasion of human triple-negative breast cancer MDA-MB-231 cells.