ALG3通过JAK/STAT通路促进三阴性乳腺癌增殖、侵袭和迁移  

ALG3 promotes proliferation, invasion and migration of triple-negative breast cancer through JAK/STAT pathway

作　　者：邢现菲 李瑞华 韩亚娟 张竞竞[1] 邵渝欢 马赛菲 刘亚萌 韩正全[1] XING Xianfei;LI Ruihua;HAN Yajuan;ZHANG Jingjing;SHAO Yuhuan;MA Saifei;LIU Yameng;HAN Zhengquan(Department of Medical Oncology,First Affiliated Hospital of Bengbu Medical College,Bengbu 233000,China;Department of Gastroenterology,First Affiliated Hospital of Bengbu Medical College;Department of Otolaryngology,Head and Neck Surgery,First Affiliated Hospital of Bengbu Medical College)

机构地区：[1]蚌埠医学院第一附属医院肿瘤内科,蚌埠233000 [2]蚌埠医学院第一附属医院消化内科 [3]蚌埠医学院第一附属医院耳鼻咽喉头颈外科

出　　处：《山西医科大学学报》2023年第5期580-588,共9页Journal of Shanxi Medical University

基　　金：安徽省教育厅自然科学研究重点项目(KJ2018A0990);蚌埠医学院自然重点项目(2020byzd156);蚌埠医学院研究生科研创新计划项目(Byycx21094)。

摘　　要：目的 探讨ALG3对乳腺癌细胞增殖、侵袭和迁移以及JAK/STAT信号通路的影响。方法 Western blot检测正常人乳腺上皮细胞HBL-100和乳腺癌细胞株SKBr3、MCF-7、MDA-MB-231中ALG3的表达情况。Western blot检测过表达和敲减序列的转染效率。先将实验分为control组(Lip 2000)、NC-KD组(Lip 2000+si-NC)和ALG3-KD组(Lip 2000+ALG3-siRNA),CCK-8,集落克隆和体内裸鼠成瘤实验检测乳腺癌细胞增殖能力;划痕实验和Transwell实验检测乳腺癌细胞侵袭和迁移能力;Western blot检测JAK/STAT通路相关蛋白JAK2/p-JAK2、STAT3/p-STAT3、MMP2、Cyclin D1、Bax表达情况。再将实验分为NC-OE组(Lip 2000+mimic-NC)、ALG3-OE组(Lip 2000+ALG3-mimic)和ALG3-OE+WP1066组(Lip 2000+ALG3-mimic+通路抑制剂WP1066),Western blot检测WP1066对JAK/STAT通路相关蛋白JAK2/p-JAK2、STAT3/p-STAT3的影响,集落克隆和Transwell实验检测抑制该通路后,ALG3对乳腺癌细胞增殖和侵袭迁移能力的影响。结果 与正常人乳腺上皮细胞HBL-100相比,ALG3在SKBr3、MCF-7、MDA-MB-231细胞中的表达较高,以MDA-MB-231细胞中的升高水平最显著(P<0.05),用于后续实验。与control组和NC-KD/OE组相比,ALG3-S3组ALG3敲减效率最佳,ALG3-mimic组过表达差异具有统计学意义(P<0.01),故用于后续敲减和过表达。与control组和NC-KD组相比,ALG3-KD组MDA-MB-231细胞增殖和侵袭迁移能力受到抑制(P<0.05),JAK/STAT通路相关蛋白JAK2/p-JAK2、STAT3/p-STAT3、MMP2、Cyclin D1的表达水平下降,凋亡相关蛋白Bax升高(P<0.05)。与NC-OE和ALG3-OE组相比,ALG3-OE+WP1066组JAK/STAT通路相关蛋白JAK2/p-JAK2、STAT3/p-STAT3及其磷酸化受到抑制(P<0.05),同时,乳腺癌细胞的增殖和侵袭迁移能力也被抑制(P<0.05)。结论 ALG3可能通过激活JAK/STAT通路促进三阴性乳腺癌细胞的增殖、侵袭和迁移。Objective To investigate the effects of ALG3 on the proliferation,invasion and migration abilities of breast cancer cells and JAK/STAT signalling pathway.Methods Western blot was performed to detect the expression of ALG3 in normal human breast epithelial cells HBL-100 and breast cancer cell lines SKBr3,MCF-7 and MDA-MB-231.Western blot was performed to detect the transfection efficiency of overexpression and knockdown sequences.The experiment was divided into control group(Lip 2000),NC-KD group(Lip 2000+si-NC)and ALG3-KD group(Lip 2000+ALG3-siRNA).CCK-8,colony cloning and in vivo nude mouse tumorigenesis assay were used to detect the proliferation ability of breast cancer cells,scratch assay and Transwell assay were used to detect the cell invasion,and Western blot was performed to detect the expression of JAK2/p-JAK2,STAT3/p-STAT3,MMP2,Cyclin D1 and Bax.The experiment was divided into NC-OE group(Lip 2000+mimic-NC),ALG3-OE group(Lip 2000+ALG3-mimic)and ALG3-OE+WP1066 group(Lip 2000+ALG3-mimic+pathway inhibitor WP1066).The effect of WP1066 on JAK2/p-JAK2 and STAT3/p-STAT3 proteins was examined by Western blot,and the effect of ALG3 on proliferation,invasion and migration abilities of breast cancer cells after inhibiting the pathway was examined by colony formation assay and Transwell assay.Results Compared with normal human breast epithelial cells HBL-100,ALG3 expression was increased in SKBr3,MCF-7 and MDA-MB-231 cells,and it was the highest in MDA-MB-231 cells(P<0.05),which were used for subsequent experiments.Compared with control group and NC-KD group,ALG3 knockdown efficiency was the best in ALG3-S3 group,and the overexpression in ALG3-mimic group was successfully established(P<0.01).Compared with control group and NC-KD group,the proliferation,invasion and migration abilities were inhibited in ALG3-KD group(P<0.05),the expression levels of JAK2/p-JAK2,STAT3/p-STAT3,MMP2,Cyclin D1 were decreased,and the expression of apoptosis-related protein Bax was increased(P<0.05).Compared with NC-OE group and ALG3-OE g

关 键 词：ALG3 JAK/STAT通路 三阴性乳腺癌 MDA-MB-231细胞 裸鼠 

分 类 号：R737.9[医药卫生—肿瘤]