摘要
目的探讨微小RNA-769-3p(miR-769-3p)对卡波氏肉瘤相关疱疹病毒(KSHV)感染的神经细胞SH-SY5Y(SK-RG)增殖及迁移方面的作用。方法将SK-RG细胞分为NC mimics转染组(转染NC mimics)、miRNA-769-3p mimics转染组(转染miRNA-769-3p mimics)、miRNA-6815-5p mimics转染组(转染miRNA-6815-5p mimics)、miRNA-6874-3p mimics转染组(转染miRNA-6874-3p mimics)和miRNA-7974 mimics转染组(转染miRNA-7974 mimics)。用实时荧光定量逆转录聚合酶链反应和蛋白质印迹法检测哺乳动物西罗莫司靶蛋白(mTOR)mRNA和蛋白的表达水平,用双荧光素酶报告基因实验检测miR-769-3p对mTOR的靶向调控,用噻唑蓝法和平板克隆实验检测细胞的增殖情况,用划痕实验和Transwell实验检测细胞的迁移情况。结果SK-RG细胞中NC mimics转染组、miRNA-769-3p mimics转染组的mTOR蛋白相对表达水平分别为1.00±0.06和0.60±0.04,细胞第5天的增殖率为4.15±0.14和3.36±0.17,克隆形成数分别为(82.67±3.51)和(53.00±3.61)个,相对迁移率分别为1.00±0.01和0.66±0.01,迁移细胞数分别为(174.30±5.86)和(131.00±5.29)个,miRNA-769-3p mimics转染组的上述指标与NC mimics转染组比较,差异均有统计学意义(均P<0.05)。结论miR-769-3p可能通过靶向抑制mTOR的表达而降低SK-RG细胞的增殖和迁移能力,提示miR-769-3p可能作为治疗KSHV感染的小分子核酸药物。
Objective To investigate the effect of microRNA-769-3p(miR-769-3p)on the proliferation and migration of SH-SY5Y(SK-RG)cells infected with Kaposi’s sarcoma-associated herpesvirus(KSHV).Methods SK-RG cells were divided into NC mimics transfection group(transfected with NC mimics),miRNA-769-3p mimics transfection group(transfected with miRNA-769-3p mimics),miRNA-6815-5p mimics transfection group(transfected with miRNA-6815-5p mimics),miRNA-6874-3p mimics transfection group(transfected with miRNA-6874-3p mimics)and miRNA-7974mimics transfection group(transfected with miRNA-7974 mimics).Real-time quantitative reverse transcription polymerase chain reaction and Western blot were used to detect the expression of mammalian target of rapamycin(m TOR)mRNA and protein.Dual luciferase reporter gene assay was used to detect the targeting regulation of miR-769-3p on m TOR.Cell proliferation was detected by methyl thiazolyl tetrazolium(MTT)assay and plate cloning assay.Cell migration was detected by scratch assay and Transwell assay.Result The relative expression levels of m TOR protein in NC mimics transfection group and miRNA-769-3p mimics transfection group were1.00±0.06 and 0.60±0.04,respectively;the proliferation rates of SK-RG cells on the 5^(th)day were 4.15±0.14and 3.36±0.17,respectively;the number of clone formation was 82.67±3.51 and 53.00±3.61,respectively;the relative migration rates were 1.00±0.01 and 0.66±0.01,respectively;the number of migrated cells was174.30±5.86 and 131.00±5.29,respectively.There were statistically significant differences in the above indicators between the miRNA-769-3p mimics transfection group and the NC mimics transfection group(all P<0.05).Conclusion MiR-769-3p may reduce the proliferation and migration ability of KSHV-infected nerve cells through targeted inhibition of m TOR expression.MiR-769-3p may be used as a nucleic acid drug to reverse the disease caused by KSHV infection.
作者
吴肇滏
丁雨飞
阿依妮萨罕·如则
潘泽民
曹冬冬
梁伟华
蒋金芳
郑志红
李冬妹
WU Zhao-fu;DING Yu-fei;AYNISAHAN Ru-ze;PAN Ze-min;CAO Dong-dong;LIANG Wei-hua;JIANG Jin-fang;ZHENG Zhi-hong;LI Dong-mei(Department of Biochemistry School of Medicine Shihezi University/Key Laboratory of Xinjiang Endemicand Ethnic Diseases Ministry of Education,Shihezi 832002,Xinjiang Uygur Autonomous Region,China;Department of Pathology,Xinjiang Production and Construction Corps Fourth Division HospitalY,ining 835099,XinjianggUygurAutonomous Region,China;Department of Pathology,Bazhou Hospital,Korla 841099,Xinjiang Uygur Autonomous Region,China)
出处
《中国临床药理学杂志》
CAS
CSCD
北大核心
2023年第19期2775-2779,共5页
The Chinese Journal of Clinical Pharmacology
基金
国家自然科学基金资助项目(81760362)
新疆生产建设兵团科技合作基金资助项目(2022BC002)
石河子大学国际合作基金资助项目(GJHZ202102)。
