文章摘要
鲍 军,焦德松,周 杨,蒋宾宾,程羽青,周晓莉.SPAG5通过影响JAK2/STAT3信号通路磷酸化促进乳腺癌细胞的增殖和侵袭[J].南京医科大学学报,2020,(4):502~508
SPAG5通过影响JAK2/STAT3信号通路磷酸化促进乳腺癌细胞的增殖和侵袭
SPAG5 promotes the proliferation and invasion of breast cancer cells through affecting the phosphorylation of JAK2/STAT3 signaling pathway
投稿时间:2020-01-20  
DOI:10.7655/NYDXBNS20200408
中文关键词: SPAG5  乳腺癌  增殖  侵袭  JAK2/STAT3
英文关键词: SPAG5  breast cancer  proliferation  invasion  JAK2/STAT3
基金项目:常州市卫生健康委青年人才科技项目(QN201935);南京医科大学科技发展基金(NMUB2019313)
作者单位
鲍 军 温岭市中医院普外二科浙江 温岭 317500 
焦德松 温岭市中医院普外二科浙江 温岭 317500 
周 杨 南京医科大学附属常州第二人民医院病理科江苏 常州 213000 
蒋宾宾 温岭市中医院普外二科浙江 温岭 317500 
程羽青 南京医科大学附属常州第二人民医院病理科江苏 常州 213000 
周晓莉 南京医科大学附属常州第二人民医院病理科江苏 常州 213000 
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中文摘要:
      目的:探究人类精子相关抗原5(sperm?associated antigen 5,SPAG5)对乳腺癌细胞增殖和侵袭的影响及分子机制。方法:采用qRT?PCR和Western blot筛选相应SPAG5高表达细胞株和低表达细胞株;分别设计并构建重组SPAG5敲减/过表达质粒(siRNA?SPAG5/OE?SPAG5),通过慢病毒表达质粒系统,制备敲减/过表达SPAG5的稳定细胞株;采用CCK?8增殖实验和裸鼠成瘤实验,检测乳腺癌细胞增殖能力的变化;采用细胞划痕实验、Transwell细胞实验检测乳腺癌细胞迁移、侵袭能力的变化;采用qRT?PCR和Western blot检测JAK2/STAT3通路和下游蛋白的变化;采用WP1066抑制JAK2/STAT3通路,检测细胞增殖和侵袭能力的变化。结果:相较于各自对照组,SPAG5敲减的 si1?MDA?MB?231组细胞SPAG5 mRNA 和蛋白水平显著降低,SPAG5过表达的OE?MCF?12A组细胞SPAG5 mRNA 和蛋白水平显著升高(P<0.05);同时,si1?MDA?MB?231组细胞的增殖、迁移、侵袭能力受到抑制,OE?MCF?12A组细胞得到增强(P<0.05);si1?MDA?MB?231组细胞的JAK2/STAT3通路磷酸化水平和下游蛋白表达受抑制(P<0.05),OE?MCF?12A组细胞的JAK2/STAT3通路磷酸化水平和下游蛋白表达水平提高(P<0.05);抑制JAK2/STAT3通路后SPAG5对乳腺癌细胞增殖和侵袭的促进作用消失(P<0.05)。结论:SPAG5通过JAK2/STAT3信号通路增强乳腺癌细胞的增殖和侵袭能力。
英文摘要:
      Objective:To investigate the effects of human sperm?associated antigen 5(SPAG5)on proliferation and invasion of breast cancer cells and its molecular mechanism. Methods:The recombinant SPAG5 knockdown/overexpression plasmids siRNA?SPAG5/OE?SPAG5 were designed and constructed respectively transfected with the lentivirus transfection system to prepare stable cell lines. CCK?8 proliferation assay and nude mouse transplantation tumor experimentwere used to detect the proliferation of breast cancer cells.Transwell experiment was used to detect the migration and invasion ability of breast cancer cells.Changes of JAK2/STAT3 pathway and downstream proteins were detected by qRT?PCR and Western blot.The JAK2/STAT3 pathway was inhibited by WP1066 to detect changes in cell proliferation and invasion. Results:The mRNA and protein levels of the SPAG5 in the si1?MDA?MB?231 group were significantly lower than those in the control group,and the mRNA and protein levels of the SPAG5 in the OE?MCF?12A were significantly higher than those in the control group(P < 0.05). Compared with the control group,the proliferation,migration and invasion ability of the si1?MDA?MB?231 group was inhibited,while these abilities in the OE?MCF?12A group were enhanced(P < 0.05). Phosphorylation level and downstream protein expression level of JAK2/STAT3 pathway in si1?MDA?MB?231 group were inhibited(P < 0.05),while phosphorylation level and downstream protein expression level of JAK2/STAT3 pathway in OE?MCF?12A group were increased(P < 0.05). Conclusion:SPAG5 promotes the proliferation and invasion of breast cancer cells through the JAK2/STAT3 signaling pathway
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