miR-195-5p靶向FGF2抑制子宫内膜癌HEC-1B细胞恶性生物学行为  

MiR-195-5p targeting FGF2 inhibits malignant biological behaviors of endometrial carcinoma HEC-1B cells

作　　者：李万斌[1] 王新勇[2] 周烨[2] LI Wanbin1, WANG Xinyong2, ZHOU Ye2(1. The Clinical College; 2. Department of Gynecology, Affiliated Hospital of Jining Medical University, Jining 272067, Shandong, China)

机构地区：[1]济宁医学院临床学院,山东济宁272067 [2]济宁医学院附属医院妇科,山东济宁272067

出　　处：《中国肿瘤生物治疗杂志》2018年第9期884-890,共7页Chinese Journal of Cancer Biotherapy

基　　金：山东省自然科学基金资助项目(No.Q98C01124)~~

摘　　要：目的:探讨miR-195-5p通过靶向FGF2抑制子宫内膜癌HEC-1B细胞增殖、凋亡、侵袭和迁移的分子机制。方法:HEC-1B细胞培养与转染完成后分为4组:HEC-1B组、miR-195-5p mimic组、p LV-FGF2组和miR-195-5p+FGF2组。q RT-PCR检测细胞miR-195-5p和FGF2 m RNA水平,荧光素酶实验验证miR-195-5p与FGF2的靶向关系,Western blotting检测FGF2表达水平,CCK-8法检测HEC-1B细胞增殖水平,流式细胞术检测HEC-1B细胞凋亡率,Transwell实验检测HEC-1B细胞侵袭能力,划痕实验检测HEC-1B细胞迁移能力。结果:与HEC-1B组相比,miR-195-5p mimic组miR-195-5p表达升高、FGF2 m RNA水平下降(均P<0.01);miR-195-5p可直接靶向FGF2。与HEC-1B组相比,miR-195-5p mimic组FGF2的蛋白表达水平下降,p LV-FGF2组FGF2的蛋白水平明显上升,且miR-195-5p+FGF2组FGF2的蛋白表达水平低于p LV-FGF2组(均P<0.01)。miR-195-5p mimic组细胞增殖水平低于HEC-1B组,p LV-FGF2组细胞增殖水平高于HEC-1B组(均P<0.01)。与HEC-1B组相比,miR-195-5p mimic组细胞凋亡率增加,p LV-FGF2组细胞凋亡率降低,且miR-195-5p+FGF2组细胞凋亡率高于p LV-FGF2组(均P<0.01)。与HEC-1B组相比,miR-195-5p mimic组每个视野下的侵袭细胞数和划痕愈合率下降,p LV-FGF2组每个视野下的侵袭细胞数和划痕愈合率上升,且miR-195-5p+FGF2组每个视野下的侵袭细胞数和划痕愈合率低于p LV-FGF2组(均P<0.01)。结论:miR-195-5p通过靶向FGF2抑制子宫内膜癌HEC-1B细胞的增殖、侵袭和迁移并促进细胞凋亡,其作为子宫内膜癌的治疗靶点。Objective： To explore the molecular mechanism of miR-195-5 p targeting FGF2 to inhibit the proliferation, apoptosis, invasion and migration of endometrial cancer HEC-1 B cells. Methods： After culture and transfection, HEC-1 B cells were divided into 4 groups： HEC-1 B group, miR-195-5 p mimic group, p LV-FGF2 group and miR-195-5 p ＋ FGF2 group. The expressions of miR-195-5 p and m RNA levels of FGF2 were detected by q RT-PCR. The targeted relationship of miR-195-5 p and FGF2 was verified by luciferase assay. The protein expression of FGF2 was examined by Western blotting; Proliferation of HEC-1 B cells was measured by CCK-8;Apoptosis was tested by flow cytometry; HEC-1 B cell invasion was detected by transwell, and migration was measured by scratch assay. Results： Compared with HEC-1 B group, the expression of miR-195-5 p in miR-195-5 p mimic group was elevated while FGF2 m RNA level was declined（all P〈0.01）. Luciferase assay indicated that FGF2 was a target of miR-195-5 p. Compared with HEC-1 B group, the protein level of FGF2 in miR-195-5 p mimic group was decreased, and the protein levels of FGF2 in p LV-FGF2 group were enhanced（P〈0.01）. The protein levels of FGF2 in miR-195-5 p＋FGF2 group were lower than that in p LV-FGF2 group（all P〈0.01）. The proliferation in miR-195-5 p mimic group was lower than HEC-1 B group（P〈0.01）, while the proliferation in p LV-FGF2 group was higher than that in HEC-1 B group（all P〈0.01）. Compared with HEC-1 B group, apoptosis in miR-195-5 p mimic group was increased,and apoptosis in p LV-FGF2 group was decreased（P〈0.01）; moreover, apoptosis in miR-195-5 p ＋ FGF2 group was higher than that in p LV-FGF2 group（P〈0.01）. Compared with HEC-1 B group, the number of invasive cells per field and the rate of wound healing in miR-195-5 p mimic group were decreased, while those in p LV-FGF2 group was enhanced（P〈0.01）; moreover, the number of invasive cells per field and the rate of wound healing in miR-195-5 p＋FGF2 group was lower t

关 键 词：子宫内膜癌 HEC-1B细胞 miR-195-5p 增殖 凋亡 侵袭 迁移 

分 类 号：R392.11[医药卫生—免疫学] R737.33[医药卫生—基础医学]