摘要
目的揭示结肠癌转移相关基因1(MACC1)对宫颈癌Hela细胞侵袭、迁移、凋亡的影响及作用机制。方法通过生物信息学分析、免疫组织化学和蛋白印迹法法检测宫颈癌组织及其对应癌旁组织中MACC1的表达。用siRNA法敲低MACC1后,通过Transwell小室、划痕、流式细胞术实验评估其对细胞迁移、侵袭、凋亡的影响。结果MACC1蛋白在宫颈癌组织中的表达显著高于癌旁组织。免疫组织化学法测得MACC1在宫颈癌组织中的阳性率是其癌旁组织的5.2倍(P<0.001)。利用siRNA转染敲低Hela细胞MACC1的效率可达90%以上,敲低MACC1表达可显著抑制细胞的迁移和侵袭能力(1∶0.4和1∶0.33,均P<0.001)。敲低MACC1表达后细胞凋亡率显著上升(5.57∶19.2,P<0.001),凋亡相关蛋白caspase3和Bax的表达水平增加(1∶2.45和1∶2.89,均P<0.001),Akt和Rac1的活性显著降低(1∶0.45和1∶0.56,均P<0.01)。利用抑制剂MK-2206和EHop-016分别抑制Akt和Rac1后,Hela细胞的侵袭和迁移能力均被显著抑制(1∶0.58∶0.62和1∶0.67∶0.52,均P<0.01)。结论宫颈癌细胞中高表达MACC1,MACC1通过激活Akt/Rac1信号通路诱导宫颈癌细胞的恶性表型。
Objective To reveal the effects and mechanism of metastasis-associated colon cancer 1(MACC1)on invasion,migration and apoptosis of cervical cancer Hela cell line.Methods Expressions of MACC1 in cervical cancer and para-tumor tissues were detected by bioinformatics analysis,immunohistochemistry and Western blot.After knockdown of MACC1 by siRNA,the cell migration,invasion,and apoptosis of cervical cancer cells were assessed by transwell,wound healing,and flow cytometry assays.Results MACC1 protein was found more highly expressed in tumor tissues than in para-tumor tissues.The immunohistochemistry results showed the positive rate of MACC1 in cervical cancer tissue was 5.2 times higher than that in para-tumor tissues(P<0.001).The efficiency of knocking-down MACC1 of Hela cells by siRNA transfection could reach more than 90%,and knocking-down of MACC1 significantly inhibited migration and invasion ability of Hela cells(1 vs 0.4 and 1 vs 0.33,all P<0.001).After knockdown of MACC1,cell apoptosis rate increased significantly(5.57 vs 19.2,P<0.001).Expression levels of apoptosis-associated proteins caspase3 and Bax increased dramatically(1 vs 2.45 and 1 vs 2.89,all P<0.001).The activities of Akt and Rac1 were significantly decreased(1 vs 0.45 and 1 vs 0.56,all P<0.01).The MK-2206 and EHop-016 were used to inhibit Akt and Rac1,respectively,and it demonstrated that the invasion and migration ability of Hela cells was significantly inhibited(1 vs 0.58 vs 0.62 and 1 vs 0.67 vs 0.52,all P<0.01).Conclusions MACC1 is highly expressed in cervical cancer cells.MACC1 could induce malignant phenotypes in cervical cancer cells by activating the Akt/Rac1 signaling pathway.
作者
王建凯
闫春梅
王文娟
马晓春
缪国英
Wang Jiankai;Yan Chunmei;Wang Wenjuan;Ma Xiaochun;Miao Guoying(Department of Radiation Oncology,Gansu Provincial Hospital,Lanzhou 730000,China;epartment of Obstetrics and Gynecology,Lanzhou Jiaotong University Hospital,Lanzhou 730070,China;Physical Examination Center,Gansu Third People′s Hospital,Lanzhou 730000,China;Department of Radiation Oncology,Gansu Provincial Central Hospital,Lanzhou 730050,China)
出处
《中华转移性肿瘤杂志》
2023年第4期410-416,共7页
Chinese Journal of Metastatic Cancer
基金
甘肃省自然科学基金(21JR11RA197,21JR1RA033)
甘肃省人民医院重点项目(ZX-62000001-2021-266)。
