摘要
目的探讨miR-208a-3p靶向亮氨酸拉链转录因子样1(LZTFL1)对宫颈癌细胞增殖、侵袭转移的影响和分子机制。方法收集2016年5月~2018年3月于湖北省中西医结合医院妇产科收治并经病理科确诊的40例宫颈癌患者手术标本。实时荧光定量PCR(RT-qPCR)和蛋白质印记(Western Blot)检测宫颈癌组织和与其对应的癌旁组织中miR-208a-3p和LZTFL1的表达。将将宫颈癌细胞Hela分为anti-miR-NC组(转染anti-miR-NC)、anti-miR-208a-3p组(转染anti-miR-208a-3p)、pcDNA组(转染pcDNA)、pcDNA-LZTFL1组(转染pcDNA-LZTFL1)。采用MTT法检测细胞活力,Transwell实验检测细胞迁移和侵袭数目,western blot检测细胞周期素D1(CyclinD1)、基质金属蛋白酶2(MMP-2)和MMP-9蛋白的表达。结果相较于癌旁组织,宫颈癌组织中miR-208a-3p的表达显著升高,LZTFL1的表达显著降低(P<0.05)。miR-208a-3p靶向LZTFL1并负性调控其表达。与anti-miR-NC组比较,anti-miR-208a-3p组Hela细胞活力、迁移侵袭能力、CyclinD1、MMP-2和MMP-9蛋白的表达显著降低,p21的表达显著升高,差异均有统计学意义(P<0.05)。与pcDNA组比较,pcDNA-LZTFL1组Hela细胞活力、迁移侵袭能力、CyclinD1、MMP-2和MMP-9蛋白表达显著降低,p21表达显著升高,差异均有统计学意义(P<0.05)。与anti-miR-208a-3p+si-NC组比较,anti-miR-208a-3p+si-LZTFL1组Hela细胞活力、迁移侵袭能力、CyclinD1、MMP-2和MMP-9蛋白的表达显著升高,p21的表达显著降低,差异均有统计学意义(P<0.05)。结论宫颈癌中miR-208a-3p呈高表达,LZTFL1呈低表达。抑制miR-208a-3p通过上调LZTFL1可抑制宫颈癌细胞的增殖、迁移和侵袭。
Objective To investigate the effect of miR-208a-3p targeting LZTFL1 on the proliferation,invasion and metastasis of cervical cancer cells and its molecular mechanism.Methods From May 2016 to March 2018,surgical specimens of 40 patients with cervical cancer were collected from Department of gynecology and obstetrics,Hubei provincial hospital of integrated traditional Chinese&Western medicine.The expressions of miR-208a-3p and LZTFL1 in cervical cancer tissues and their adjacent tissues were detected by real-time fluorescence quantitative PCR(RT-qPCR)and Western Blot.StarBase online prediction,luciferase reporter gene experiments and Western Blot were used to verify the targeting relationship between miR-208a-3p and LZTFL1.Hela cells were divided into anti-miR-NC,anti-miR-208a-3p,pcDNA,pcDNA-LZTFL1,anti-miR-208a-3p+si-NC,anti-miR-208a-3p+si-LZTFL1 groups.Cell viability was measured by MTT method.The number of cell migration and invasion was measured by the Transwell test.The expression of CyclinD1,MMP-2 and MMP-9 proteins was detected by Western blot.Results Compared with adjacent cancer tissues,the expression of miR-208a-3p was significantly increased in cervical cancer tissues,while the expression of LZTFL1 was significantly decreased(P<0.05).miR-208a-3p targets LZTFL1 and negatively regulates its expression.Compared with the anti-miR-NC group,cell viability,migration and invasion ability,expression of CyclinD1,MMP-2 and MMP-9 protein of Hela cell in anti-miR-208a-3p group was significantly reduced,whereas the expression of p21 was significantly increased(P<0.05).Compared with the pcDNA group,cell viability,migration and invasion ability,expression of CyclinD1,MMP-2 and MMP-9 protein of Hela cell in pcDNA-LZTFL1 group was significantly reduced,whereas the expression of p21 was significantly increased,the difference was statistically significant(P<0.05).Compared with anti-miR-208a-3p+si-NC group,cell viability,migration and invasion ability,expression of CyclinD1,MMP-2 and MMP-9 proteins of Hela cell in anti-miR-2
作者
李利玲
王少帅
汪向红
徐铮铮
肖书瑶
LI Liling;WANG Shaoshuai;WANG Xianghong;XU Zhengzheng;XIAO Shuyao(Department of Obstetrics and Gynecology,Hubei Provincial Hospital of Integrated Chinese&Western Medicine,Wuhan 430015,China;Department of Obstetrics and Gynecology,Tongji Hospital,Tongji Medical College,Huazhong University of Science and Technology,Wuhan 430030,China)
出处
《西部医学》
2020年第9期1292-1297,共6页
Medical Journal of West China
