干扰LINC01133对宫颈癌细胞增殖、炎性因子水平、氧化应激以及裸鼠成瘤的影响  

Effect of Interfering with LINC01133 on Proliferation,Inflammatory Factors Secretion,Oxidative Stress in Cervical Cancer Cells,and Tumorigenesis in Nude Mice

作　　者：李芬[1] 古丽比亚·艾则孜 苑锦睿 亚力坤·穆罕穆德 温蒙科 沈谷群[1] LI Fen;Guliya·Aizezi;YUAN Jinrui;Yalikun·Muhanmude;WEN Mengke;SHEN Guqun(Department of Gynecology and Surgery,Cancer Hospital Affiliated to Xinjiang Medical University,Urumqi,830011,China)

机构地区：[1]新疆医科大学附属肿瘤医院妇外二科,乌鲁木齐市830011

出　　处：《医学分子生物学杂志》2023年第4期292-297,共6页Journal of Medical Molecular Biology

基　　金：新疆维吾尔自治区自然科学基金(No.2020D01C204);新疆维吾尔自治区科技支疆项目(No.2020E02125)。

摘　　要：目的探究干扰长链非编码RNA LINC01133在宫颈癌细胞中的作用和机制。方法TCGA数据库和qRT-PCR分析LINC01133在宫颈癌组织和细胞中的表达;qRT-PCR检测LINC01133-shRNAs的敲低效率;SiHa细胞分为Control组、shRNA-NC组、LINC01133-shRNA1组,克隆形成实验检测细胞增殖,ELISA检测肿瘤坏死因子-α(TNF-α)、白细胞介素(IL)-6和IL-1β水平;免疫荧光检测SiHa细胞中P65的核转移;Western印迹检测P65的磷酸化;试剂盒检测超氧化物歧化酶(SOD)和乳酸脱氢酶(LDH);建立裸鼠移植瘤模型,分析干扰LINC01133表达对肿瘤生长的影响。结果LINC01133在宫颈癌组织和细胞中的表达显著上调(P<0.05)。LINC01133-shRNA1组敲除效率最为显著,选择此组作为后续实验干扰组;与对照组相比,LINC01133-shRNA1组克隆形成率、炎性因子水平、P65磷酸化水平、细胞核P65含量和LDH水平均显著升高(P<0.05),SOD活性显著降低(P<0.05);在裸鼠成瘤实验中,与shRNA-NC组相比,LINC01133-shRNA1组肿瘤组织体积、重量、SOD活性均显著降低(P<0.05),P65的磷酸化水平显著升高(P<0.05)。结论干扰LINC01133抑制宫颈癌细胞生长、炎性因子水平和氧化应激。Objective To explore the function and mechanism of interfering long non-coding RNA LINC01133 in cervical cancer cells.Methods TCGA database and qRT-PCR were used to analyze the expression of LINC01133 in cervical cancer tissues and cells.The knockdown efficiency of LINC01133-shRNAs was detected by qRT-PCR.SiHa cells were divided into 3 groups:control group,shRNA-NC group and LINC01133-shRNA1 group.Colony formation assay was used to detect cell proliferation.ELISA assay was used to detect the levels of tumor necrosis factor-α(TNF-α),interleukin(IL)-6 and IL-1β.The nuclear translocation of P65 in SiHa cells was detected by immunofluorescence.The phosphorylation of P65 was detected by Western blotting.The levels of superoxide dismutase(SOD)and lactate dehydrogenase(LDH)were detected by kits.A nude mouse xenograft model was established to analyze the effect of interfering LINC01133 expression on tumor growth.Results The expression of LINC01133 was significantly up-regulated in cervical cancer tissues and cells(P<0.05).The knockdown efficiency of LINC01133-shRNA1 was the best,thus the LINC01133-shRNA1 was selected for the subsequent experiments.The colony formation rate,the levels of inflammatory factors,the phosphorylation level of P65,the proportion of P65 level in nuclear,and the LDH level in the LINC01133-shRNA1 group were significantly increased when compared with those in the control group(P<0.05),while the SOD activity was significantly decreased(P<0.05).The volume weight of the tumor tissues,and the SOD activity in the LINC01133-shRNA1 group were significantly decreased when compared with those in the shRNA-NC group(P<0.05),while the phosphorylation level of P65 was significantly increased(P<0.05).Conclusion Interfering with the LINC01133 shRNA inhibits cervical cancer cell growth,decreases inflammatory factor levels and suppresses the oxidative stress in cervical cancer cells.

关 键 词：宫颈癌 LINC01133 炎性因子 超氧化物歧化酶 P65 

分 类 号：R737.33[医药卫生—肿瘤]