信号转导和转录激活因子3/CC趋化因子配体2信号通路对乳腺癌细胞活力、迁移和侵袭的影响实验研究OACSTPCD
Effect of signal transducer and activator of transcription 3/C-C motif chemokine ligand 2 signaling pathway on viability,migration and invasion of breast cancer cells
目的:探讨信号转导和转录激活因子3(STAT3)/CC趋化因子配体2(CCL2)信号通路对乳腺癌细胞活力、迁移和侵袭的影响.方法:体外培养人乳腺癌细胞株(HCC1937、MCF-7、MDA-MB-231、ZR-75-1)和人正常乳腺上皮细胞(MCF-10A).采用蛋白印迹法检测HCC1937、MCF-7、MDA-MB-231、ZR-75-1和MCF-10A细胞磷酸化STAT3(p-STAT3)、STAT3、CCL2蛋白表达.选取p-STAT3/STAT3蛋白比值、CCL2蛋白表达最高的人乳腺癌细胞进行后续实验.将MCF-7细胞分为对照组、STAT3抑制剂(WP1066)Ⅰ组(2 μmol/L WP1066)、Ⅱ组(4 μmol/L WP1066)、Ⅲ组(8 μmol/L WP1066).分别采用细胞计数试剂盒-8、划痕实验、Transwell实验检测MCF-7细胞活力、迁移率及侵袭能力.采用蛋白印迹法检测MCF-7细胞p-STAT3、STAT3、CCL2、基质金属蛋白酶(MMP)-2、MMP-9 蛋白表达.结果:与 MCF-10A 细胞比较,HCC1937、MCF-7、MDA-MB-231、ZR-75-1 细胞p-STAT3/STAT3蛋白比值、CCL2蛋白表达升高(均P＜0.05).其中MCF-7细胞p-STAT3/STAT3蛋白比值、CCL2蛋白表达最高,因此选用MCF-7细胞进行后续实验.与对照组比较,STAT3抑制剂Ⅰ、Ⅱ、Ⅲ组MCF-7细胞活力、迁移率、侵袭数目依次降低(均P＜0.05).与对照组比较,STAT 3抑制剂Ⅰ、Ⅱ、Ⅲ组MCF-7细胞p-STAT 3/STAT 3蛋白比值、CCL2、MMP-2及MMP-9蛋白表达依次降低(均P＜0.05).结论:乳腺癌细胞中STAT3/CCL2呈高表达,抑制STAT3/CCL2信号通路能够降低乳腺癌细胞活力,减少迁移和侵袭.
Objective:To investigate the effect of signal transducer and activator of transcription 3(STAT 3)/C-C motif chemokine ligand 2(CCL2)signaling pathway on viability,migration and invasion of breast cancer cells.Methods:Human breast cancer cell lines(HCC1937,MCF-7,MDA-MB-231,ZR-71-1)and human normal breast epithelial cells(MCF-10A)were cultured in vitro.The expressions of p-STAT3,STAT3 and CCL2 in HCC1937,MCF-7,MDA-MB-231,ZR-75-1 and MCF-10A cells were detected by Western blot.Human breast cancer cells with the highest p-STAT3/STAT3 protein ratio and CCL2 protein expression were selected for follow-up experiments.MCF-7 cells were divided into control group,STAT3 inhibitor(WP1066)group Ⅰ(2 μmol/L WP1066),group Ⅱ(4 μmol/L WP1066)and groupⅢ(8 μmol/L WP1066).Cell count kit 8,scratch test and Transwell test were used to detect MCF-7 cell viability,mobility and invasion ability,respectively.The protein expressions of p-STAT3,STAT3,CCL2,matrix metalloprotei-nase(MMP)-2 and MMP-9 in MCF-7 cells were detected by Western blot.Results:Compared with MCF-10A cells,the expressions of p-STAT3/STAT3 protein ratio and CCL2 protein were increased in HCC1937,MCF-7,MDA-MB-231 and ZR-75-1 cells(all P＜0.05).Among them,the p-STAT3/STAT3 protein ratio and CCL2 protein expression were the highest in MCF-7 cells,so MCF-7 cells were selected for follow-up experiments.Compared with the control group,MCF-7 cells viability,mobility and invasion number in STAT3 inhibitor groups Ⅰ,Ⅱ and Ⅲ were decreased successively(all P＜0.05).Compared with the control group,the p-STAT3/STAT3 protein ratio,CCL2,MMP-2 and MMP-9 protein expressions of MCF-7 cells in STAT3 inhibitor groups Ⅰ,Ⅱ and Ⅲ were decreased successively(all P＜0.05).Conclusion:STAT3/CCL2 is highly expressed in breast cancer cells,and inhibition of STAT3/CCL2 signaling pathway can reduce the viability,migration and invasion of breast cancer cells.
薛杰;张永梅;陈启;郝艳萍
内蒙古医科大学附属医院,内蒙古呼和浩特 010050宁夏医科大学第三临床医学院,宁夏银川 750004
基础医学
乳腺癌信号转导和转录激活因子3CC趋化因子配体2细胞活力细胞迁移细胞侵袭
Breast cancerSignal transducer and activator of transcription 3C-C motif chemokine ligand 2Cell viabilityCell migrationCell invasion
《陕西医学杂志》 2024 (005)
579-582,588 / 5
内蒙古自治区自然科学基金资助项目(2019MS08150)
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