高压氧通过下调miR-720表达抑制卵巢癌SKOV-3细胞恶性生物学行为的研究  

Hyperbaric oxygen inhibits malignant biological behavior of SKOV-3 ovarian cancer cells through down-regulation of miR-720 expression

作　　者：杨冬梅[1] 殷桂云[1] 张晓飞 Yang Dongmei;Yin Guiyun;Zhang Xiaofei(Department of Gynelology,Linyi People’s Hospital,Linyi 276000,China)

机构地区：[1]临沂市人民医院妇科,临沂276000

出　　处：《中华航海医学与高气压医学杂志》2023年第4期424-428,共5页Chinese Journal of Nautical Medicine and Hyperbaric Medicine

摘　　要：目的探讨高压氧(HBO)处理对卵巢癌SKOV-3细胞miR-720的表达及其恶性生物学行为的影响。方法卵巢癌SKOV-3细胞按照实验设计分为对照组和HBO组,对照组SKOV-3细胞常规培养,HBO组SKOV-3细胞常规培养完成后予以0.2 MPa HBO处理。同时将培养好的卵巢癌SKOV-3细胞按照随机数字表法再分为3组,正常对照(NC)组、HBO组和HBO+miR-720组,其中HBO组按照HBO条件进行培养,HBO+miR-720组在HBO处理前转染miR-720质粒,HBO组和NC组同时转染NC质粒,转染24 h后收集细胞进行后续实验。应用qPCR检测SKOV-3细胞miR-720 mRNA表达水平,应用Western印迹法检测SKOV-3细胞上皮-间充质转化(EMT)相关蛋白(N-cadherin、E-cadherin、Vimentin)和miR-720蛋白的表达水平,应用CCK-8法检测SKOV-3细胞增殖能力,应用克隆形成实验检测SKOV-3细胞克隆形成能力,应用细胞划痕实验和Transwell实验检测SKOV-3细胞侵袭和迁移能力。结果应用HBO处理后,HBO组卵巢癌SKOV-3细胞的增殖能力、侵袭能力及克隆形成能力明显低于对照组,划痕宽度比高于对照组(P<0.01);HBO组卵巢癌SKOV-3细胞中miR-720 mRNA和蛋白表达水平均低于对照组(P<0.01)。转染后,HBO+miR-720组SKOV-3细胞中miR-720 mRNA和蛋白水平明显升高(P<0.05);HBO+miR720组SKOV-3细胞的增殖能力、侵袭能力及克隆形成能力明显高于对照组,划痕宽度比明显低于对照组(P<0.01);HBO+miR-720组SKOV-3细胞中E-cadherin蛋白表达水平低于HBO组,Vimentin和N-cadherin蛋白表达高于HBO组(P<0.01)。结论 HBO处理能够明显抑制卵巢癌SKOV-3细胞的增殖、侵袭、迁移和克隆形成,其抑制作用与卵巢癌细胞中miR-720表达有关。Objective To investigate the effect of hyperbaric oxygen(HBO)on the expression of miR-720 in SKOV-3 ovarian cancer cells and malignant biological behavior,as well as cell biology and epithelial-mesenchymal transition(EMT).Methods The SKOV-3 ovarian cancer cells were divided into control group and HBO group.The cells in the control group were conventionally cultured.The HBO group was treated with hyperbaric oxygen of 0.2 MPa after conventional culture.The ovarian cancer cells were then randomly divided into three sub-groups according to the random number method:normal control(NC)group,HBO group,and HBO+miR-720 group.The HBO group was cultured with the HBO intervention,the cells in the HBO+miR-720 group were transfected with the miR-720 plasmid before HBO intervention,and the cells in the HBO and NC groups were transfected with the NC plasmid at the same time.Twenty-four hours after transfection,the cells were collected for further tests.The miR-720 mRNA was detected by qPCR;the expressions of EMT proteins,such as N-cadherin,E-cadherin,Vimentin,and miR-720,were detected by Western blotting assay;cell proliferation ability was detected by CCK-8 assay,and clonality was detected by clonality assay;cell invasion and migration abilities were detected by cell scratch assay and Transwell assay.Results After HBO intervention,the abilities of proliferation,invasion,and clonality of SKOV-3 ovarian cancer cells of the HBO group were lower than those of the control group,while the scratch width was higher than that of the control group(P<0.01).After HBO intervention,the mRNA and protein expression levels of miR-720 in ovarian cancer cells of the HBO group were lower than those of the control group(P<0.01).After transfection,the mRNA and protein expression levels of miR-720 in the HBO+miR-720 group were obviously increased(P<0.05).The abilities of proliferation,invasion,and clonality of the HBO+miR-720 group were higher than those in the NC group,while the scratch width was significantly lowered(P<0.01).Compared with the HBO gr

关 键 词：高压氧 miR-720 卵巢癌 恶性生物学行为 

分 类 号：R737.31[医药卫生—肿瘤]