Ran Wei, Zeng Yu-Hua, Ma Xiao-Jie, Liao Ping, Liu Xing-Lan
Department of Obstetrics and Gynecology, Affiliated Hospital of North Sichuan Medical College, Nanchong 637000, China.
Department of Oncology, Affiliated Hospital of North Sichuan Medical College, Nanchong 637000, China.
Sichuan Da Xue Xue Bao Yi Xue Ban. 2020 Mar;51(2):178-184. doi: 10.12182/20200360501.
To investigate the effect of miR-503-5p on the proliferation, invasion, migration and epithelialization of cervical cancer HeLa cells via targeting 2 3.
Four ccervical cancer HeLa cells groups were set up including control group, mimic-NC group, miR-503-5p mimic group, 2 3 group, miR-503-5p mimic+ 2 3 group (mimic+ 2 3 group). The plasmids were separately or jointly transinfected into cervical cancer Hela cells of each group by Lipofectamine 2000, After transinfection, the target gene was predicted by gene prediction software, the targeting relationship was verified by fluorescein experiment, the expression of miR-503-5p and 2 3 was detected by RT-PCR, cell proliferation was detected by MTT assay, expression of Ki67, proliferating cell nuclear antigen (PCNA), E-cadherin and N-cadherin were detected by Western blot, cell invasion was detected by Transwell, and cell migration was detected by scratch test. Nude mice were divided into control group and miR-503-5p mimic group, and 0.2 mL of cervical cancer HeLa cell suspension transfected with mimic-NC or miR-503-5p mimic was injected subcutaneously into the ventral side of the right hind limb of nude mice. Thirty days post injection, the nude mice were sacrificed by cervical dislocation. The tumor weight was weighed by an electronic balance, and the expression of KI67 and Vimentin in the tumor tissue was detected by immunohistochemistry.
The expression level of miR-503-5p in cervical cancer HeLa cells was down-regulated, miR-503-5p directly targeted 2 3 by binding with 2 3 at binding sites in the 3'UTR region. Over-expressing of miR-503-5p inhibited the expression of 2 3, significantly decreased cell growth rate and the expression level of Ki67 and PCNA, decreased the number of invasive cells, widened the scratches, reduced the healing rate, up-regulated the expression of E-cadherin and also down-regulated the expression of N-cadherin ( <0.01). Over-expressing of miR-503-5p significantly reduced the volume and weight of transplanted tumors, and decreased the proportion of positive Ki67 and Vimentin ( <0.01).
miR-503-5p inhibits the proliferation, invasion, migration and epithelialization of cervical cancer HeLa cells by targeting 2 3.
通过靶向2 3研究miR - 503 - 5p对宫颈癌HeLa细胞增殖、侵袭、迁移及上皮化的影响。
设立四组宫颈癌HeLa细胞组,包括对照组、模拟物阴性对照组(mimic - NC组)、miR - 503 - 5p模拟物组、2 3组、miR - 503 - 5p模拟物 + 2 3组(模拟物 + 2 3组)。通过Lipofectamine 2000将质粒分别或联合转染至各组宫颈癌Hela细胞中。转染后,用基因预测软件预测靶基因,通过荧光素实验验证靶向关系,用RT - PCR检测miR - 503 - 5p和2 3的表达,用MTT法检测细胞增殖,用蛋白质免疫印迹法检测Ki67(细胞增殖相关抗原)、增殖细胞核抗原(PCNA)、E - 钙黏蛋白和N - 钙黏蛋白的表达,用Transwell检测细胞侵袭,用划痕实验检测细胞迁移。将裸鼠分为对照组和miR - 503 - 5p模拟物组,将转染了模拟物阴性对照或miR - 503 - 5p模拟物的0.2 mL宫颈癌HeLa细胞悬液皮下注射到裸鼠右后肢腹侧。注射30天后,通过颈椎脱臼处死裸鼠。用电子天平称取肿瘤重量,用免疫组织化学法检测肿瘤组织中KI67和波形蛋白的表达。
宫颈癌HeLa细胞中miR - 503 - 5p的表达水平下调,miR - 503 - 5p通过与3'UTR区域的结合位点与2 3结合直接靶向2 3。过表达miR - 503 - 5p抑制2 3的表达,显著降低细胞生长速率以及Ki67和PCNA的表达水平,减少侵袭细胞数量,使划痕变宽,降低愈合率,上调E - 钙黏蛋白的表达并下调N - 钙黏蛋白的表达(P <0.01)。过表达miR - 503 - 5p显著减小移植瘤的体积和重量,并降低Ki67和波形蛋白阳性比例(P <0.01)。
miR - 503 - 5p通过靶向2 3抑制宫颈癌HeLa细胞的增殖、侵袭、迁移及上皮化。
需注意,原文中“2 3”表述不太清晰准确,可能影响对整体含义的精准理解,以上译文按原样翻译。