摘要
目的细胞凋亡及上皮细胞-间充质转化(EMT)是肿瘤发生、进展以及复发的重要机制之一,已成为研究肿瘤的热点。文中旨在探讨青藤碱对膀胱癌细胞株T24侵袭转移、凋亡以及EMT的影响。方法体外培养膀胱癌细胞株T24细胞,采用不同浓度青藤碱(0、0.5、1.0、2.0mmol/L)处理细胞24h后,分别采用细胞划痕实验及Transwell实验检测青藤碱对细胞侵袭及迁移的影响;同时采用MTT、流式细胞术检测细胞活性以及凋亡,Western blot检测细胞凋亡相关分子B cl-2/Bak的表达以及A kt的磷酸化,EMT相关分子表达水平;采用PI3K/Akt抑制剂处理细胞,Western blot检测EMT相关分子E-cadherin、N-cadherin、Slug和Snial的变化情况。结果MTT检测结果显示,0.5、1.0、2.0 mmol/L的青藤碱处理24h后,膀胱癌细胞株T 24的细胞活性随着青藤碱的浓度升高而逐渐降低[(100.00±2.52)%、(86.5±5.2)%、(64.5±4.5)%、(53.5±7.2)%],差异有统计学意义(P<0.05).0.5、1.0、2.0mmol/L的青藤碱处理48h后,膀胱癌细胞株T24的凋亡率显著增高。划痕实验结果显示,随着青藤碱浓度增加(0~2.0 mmol/L),膀胱癌细胞株T24的迁移水平呈逐渐下降的趋势。Transwell实验结果表明,随着青藤碱浓度增加,膀胱癌细胞株T24的侵袭率呈逐渐下降的趋势。2.0 mmol/L青藤碱处理后,膀胱癌细胞株T24中EMT相关分子E-cadherin表达上调,N-Cadherin、Slug和Snial表达下调。采用PI3K/Akt抑制剂处理细胞结果显示,抑制剂LY294002处理的膀胱癌细胞株T24中E-cadherin表达水平显著升高,而N-cadherin、Slug和Snial的表达水平显著降低。结论青藤碱能够抑制膀胱癌细胞株T24的增殖、侵袭以及迁移,并促进细胞凋亡,同时还能够通过抑制PI3K/A kt信号通路抑制膀胱癌细胞株T24发生EMT,为膀胱癌的临床治疗提供了实验依据。
Objective To investigate the effect of sinomenine(SIM) on the invasion, metastasis, apoptosis and epithelial mesenchymal transition(EMT) of T24 bladder cancer cells. Methods Bladder cancer cell line T24 was cultured in vitro and treated with SIM of different concentrations for 24 hours. Cell wound healing assay and Boyden chamber assay were used to detect the effect of SIM on cell invasion and migration. Meanwhile, MTT and flow cytometry were used to detect cell viability and apoptosis. Western blot was used to detect the expression of apoptosis related molecule Bcl-2/Bak and Akt phosphorylation. The expression levels of EMT related molecules E-cadherin, N-cadherin, slug and NiAl were measured by Western blot. After cells were treated with PI3 K/Akt inhibitors, the changes of EMT related molecules E-cadherin, N-cadherin, slug and NiAl were detected to confirm the role of PI3 K/Akt in the inhibition of EMT in T24 cells by SIM. Results Different concentrations of SIM inhibited the invasion rate and migration level of T24 cell line, reduced cell activity, and promoted cell apoptosis. Western blot showed that SIM treatment decreased bcl-2 expression and Akt phosphorylation, and increased Bak expression. In addition, the expression of E-cadherin was up-regulated, while the expression of N-cadherin, slug and NiAl were down regulated in T24 cell line treated with 2.0 mmol/l SIM. After pretreatment of T24 cells with ly2940023, an inhibitor of PI3 K/Akt, the expression of E-cadherin was higher than that of cells treated with SIM alone, while the expression of N-cadherin, slug and NiAl decreased significantly(P< 0.05).Conclusion SIM can inhibit the proliferation, invasion and migration of bladder cancer cell line T24, and promote apoptosis. SIM can also inhibit EMT of bladder cancer cell line T24 by inhibiting PI3 K/Akt signal pathway.
作者
吴磊
黎炼
罗志刚
张涛
WU Lei;LI Lian;LUO Zhi-gang;ZHANG Tao(Department of Urology,the Second Affiliated Hospital of University of South China,Hengyang 421001,Hunan,China)
出处
《医学研究生学报》
CAS
北大核心
2020年第9期909-913,共5页
Journal of Medical Postgraduates
基金
国家自然科学基金(81273754)。
关键词
青藤碱
膀胱癌
凋亡
上皮细胞-间充质转化
sinomenine
osteosarcoma cells
apoptosis
epithelial-mesenchymal transition
