TGF-β1通过激活NOX4/ROS通路促进宫颈癌细胞的侵袭和迁移  

TGF-β1 activates NOX4/ROS pathway to promote the invasion and migration of cervical cancer cells

作　　者：汪洪涛[1] 王井 刘红丽[3] 王效静[4] WANG Hongtao;WANG Jing;LIU Hongli;WANG Xiaojing(Anhui Key Laboratory of Infection and Immunity, Bengbu Medical College, Bengbu 233030;Department of Obstetrics and Gynecology, Guoyang Hospital of Traditional Chinese Medicine, Bozhou 236800;Department of Gynecological Oncology, First Afliliated Hospital, Bengbu Medical College,Bengbu 233004, China;Anhui Clinical and Preclinical Key Laboratory of Respiratory Disease, First Affiliated Hospital, Bengbu Medical College, Bengbu 233004, China)

机构地区：[1]蚌埠医学院感染与免疫安徽省重点实验室,安徽蚌埠233030 [2]涡阳县中医院妇产科,安徽亳州236800 [3]蚌埠医学院第一附属医院妇瘤科 [4]蚌埠医学院第一附属医院呼吸系病临床基础安徽省重点实验室,安徽蚌埠233004

出　　处：《细胞与分子免疫学杂志》2019年第2期121-127,共7页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家自然科学基金(81772493);安徽高校省级自然科学研究项目(KJ2016A472);蚌埠医学院科技发展基金项目(BYKF1726)

摘　　要：目的探讨转化生长因子β1/尼克酰胺腺嘌呤二核苷酸磷酸氧化酶/活性氧(TGF-β1/NOX4/ROS)信号通路在宫颈癌细胞侵袭和迁移中的作用。方法采用5 ng/mL TGF-β1处理培养的SiHa、 CaSki、 HeLa、 C4-I和C33A宫颈癌细胞4 h或8 h, 2′, 7′-二氯荧光素二乙酸酯(DCFH-DA)染色结合流式细胞术检测处理前后SiHa、 CaSki、 HeLa、 C4-I和C33A宫颈癌细胞ROS水平;转染NOX4小干扰RNA(NOX4-siRNA)或加入NOX4信号通路抑制剂二亚苯基碘氯化物(DPI), Western blot法检测宫颈癌HeLa细胞NOX4蛋白水平, Transwell^(TM)侵袭和迁移实验检测TGF-β1对宫颈癌细胞侵袭和迁移能力的影响。结果 TGF-β1显著上调HPV阳性SiHa、 CaSki、 HeLa和C4-I宫颈癌细胞NOX4蛋白表达,促进ROS产生,并增强宫颈癌细胞的侵袭和迁移性;敲低NOX4基因或使用NOX4通路抑制剂DPI可逆转这种作用。结论 TGF-β1激活NOX4/ROS信号通路增强宫颈癌细胞的侵袭和迁移能力。Objective To investigate the role of transforming growth factor β1/nicinamide adenine dinucleotide phosphate oxidase/reactive oxygen species(TGF-β1/NOX4/ROS) signaling pathway in the invasion and migration of cervical cancer cells. Methods The cultured SiHa, CaSki, HeLa, C4-I and C33A cervical cancer cells were treated with 5 ng/mL TGF-β1 for 4 hours or 8 hours. ROS levels in these cervical cancer cells were detected using 2′, 7′-dichlorofluorescein diacetate(DCFH-DA) staining combined with flow cytometry. After the transfection of NOX4 small interfering RNA(NOX4-siRNA) into HeLa cells or using of NOX4 signaling pathway inhibitor diphenylidene iodine chloride(DPI), NOX4 protein expression was measured by Western blotting, and the effect of TGF-β1 on migration and invasion ability of cervical cancer cells were analyzed by TranswellTM assay. Results TGF-β1 significantly up-regulated NOX4 protein expression, increased ROS production in HPV-positive cervical cancer cells(SiHa, CaSki, HeLa and C4-I) and promoted the migration and invasion abilities of cervical cancer cells. Silencing the NADPH oxidase gene or using the NOX4 pathway inhibitor DPI could reverse this effect. Conclusion TGF-β1 can activate NOX4/ROS signaling pathway to enhance the invasion and migration abilities of cervical cancer cells.

关 键 词：转化生长因子β1(TGF-β1) 尼克酰胺腺嘌呤二核苷酸磷酸氧化酶4(NOX4) 活性氧(ROS) 宫颈癌 侵袭性 

分 类 号：Q279[生物学—细胞生物学] R392-33[医药卫生—免疫学]