文章摘要
于利,张明,王迪.长链非编码 RNA小核仁 RNA宿主基因 7调控卵巢癌细胞迁移和侵袭的机制研究[J].安徽医药,2022,26(1):121-125.
长链非编码 RNA小核仁 RNA宿主基因 7调控卵巢癌细胞迁移和侵袭的机制研究
Study on the mechanism of lncRNA SNHG7 regulating migration and invasion of ovarian cancer cells
  
DOI:10.3969/j.issn.1009-6469.2022.01.028
中文关键词: 卵巢肿瘤  细胞运动  小核仁 RNA宿主基因 7  微小 RNA-34a
英文关键词: Ovarian neoplasms  Cell movement  Small nucleolar RNA host gene 7  microRNA-34a
基金项目:
作者单位
于利 盘锦市中心医院妇产科辽宁盘锦 124000 
张明 盘锦市中心医院妇产科辽宁盘锦 124000 
王迪 盘锦市中心医院妇产科辽宁盘锦 124000 
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中文摘要:
      目的研究长链非编码 RNA(lncRNA)小核仁 RNA宿主基因 7(SNHG7)调控卵巢癌 SK-OV-3细胞迁移和侵袭的机制。方法研究起止时间为 2019年 6—12月, SK-OV-3细胞购自美国 ATCC;卵巢癌 SK-OV-3细胞中转染 SNHG7干扰表达质粒(SNHG7 shRNA)实时定量 PCR方法测定细胞中 SNHG7表达变化,四甲基偶氮唑盐比色法( MTT)测定 SK-OV-3细胞增殖能力,用平板克隆实验,及 Transwell小室法分别检测细胞克隆形成、侵袭和迁移能力,用 Western blotting方法检测波形蛋白( Vi-mentin)、上皮钙黏蛋白( E-cadherin)表达。用 starbase生物信息软件发现 SNHG7和微小 RNA-34a(miR-34a)有互补结合位点,荧光素酶报告系统鉴定靶向关系。卵巢癌 SK-OV-3细胞中共转染 SNHG7 shRNA、miR-34a抑制剂( miR-34a inhibitor),检测细胞增殖、克隆、侵袭以及迁移能力变化。结果 Control组、 Sh-NC组、 Sh-SNHG7组细胞 A值分别为( 0.45±0.03)、(0.44±0.05)、(0.23±0.02);克隆数目分别为( 167.25±11.87)个、(169.57±12.76)个、(103.69±9.92)个;侵袭数目分别为( 135.68±10.75)个、(134.21±10.27)个、(76.33±8.20)个;迁移数目分别为( 178.69±15.46)个、(180.26±13.71)个、(99.83±8.64)个;与 Control组和 Sh-NC组比, Sh-SNHG7组细胞 A值降低,克隆数目减少,侵袭和迁移数目减少, Vimentin蛋白表达量减少, E-cadherin蛋白表达量升高,差异有统计学意义( P<0.001)。 SNHG7靶向调控 miR-34a。与 Sh-SNHG7+anti-miR-NC组比较, Sh-SNHG7+anti-miR-34a组 miR-34a水平降低, SK-OV-3细胞 A值升高[( 0.40±0.04)比( 0.24±0.03)]SK-OV-3细胞克隆形成数[( 152.16±10.35)个比(106.58±10.52)个]、侵袭数[(122.64±9.93)个比( 77.69±6.50)个]及迁移数[8.43±12.84)个比( 100.39±7.91)个]增多, Vimen-tin蛋白表达水平升高, E-cadherin蛋白表达水平下降,差异有统计学意义( P<0.001)。结论下调 lncRNA SNHG7抑制卵巢癌 (16,SK-OV-3细胞迁移和侵袭,机制与靶向调控 miR-34a有关。
英文摘要:
      Objective To study the mechanism of long non-coding RNA(lncRNA)small nucleolar RNA host gene 7(SNHG7)regulat-ing the migration and invasion of ovarian cancer SK-OV-3 cells.Methods Study started and ended from June 2019 to December 2019, SK-OV-3 cells were purchased from ATCC in the United States; ovarian cancer SK-OV-3 cells were transfected with SNHG7 in-terference expression plasmid (SNHG7 shRNA), real-time quantitative PCR method was used to determine the expression changes ofSNHG7 in cells, the tetramethylazolium salt colorimetric method (MTT) was used to determine the proliferation ability of SK-OV-3 cells, and the plate cloning experiment and Transwell chamber method were used to detect cell clone formation, invasion and migrationability, Western blotting method was used to detect the expression of Vimentin and epithelial cadherin (E-cadherin). SNHG7 and miR-34a were found to have complementary binding sites using starbase bioinformatics software, and the luciferase reporter system identi-fied the targeting relationship. SNHG7 shRNA and miR-34a inhibitor were co-transfected into ovarian cancer SK-OV-3 cells to detect changes in cell proliferation, cloning, invasion and migration ability.Results The cell A values of the Control group, Sh-NC group, and Sh-SNHG7 group were (0.45±0.03), (0.44±0.05), (0.23±0.02), respectively; the number of clones were (167.25±11.87), (169.57±12.76),and (103.69±9.92), respectively; the number of invasions were (135.68±10.75), (134.21±10.27), and (76.33±8.20), respectively; thenumber of migration was (178.69±15.46), (180.26±13.71) ) and (99.83±8.64), respectively. Compared with the Control group and theSh-NC group, the Sh-SNHG7 group cell A value was decreased, the number of clones was decreased, the number of invasion and migra-tion were decreased, the expression of Vimentin protein was decreased, and the expression of E-cadherin protein was increased, and the difference was statistically significant (P<0.001). SNHG7 targeted miR-34a. Compared with the Sh-SNHG7+anti-miR-NC group, the level of miR-34a in the Sh-SNHG7+anti-miR-34a group was decreased, and the A value of SK-OV-3 cells was increased [(0.40±0.04) vs. (0.24±0.03) )], SK-OV-3 cell clone formation number [(152.16±10.35) vs. (106.58±10.52)], invasion number [(122.6±49.93) vs. (77.69±6.50)] and migration number [(168.43±12.84) vs. (100.39±7.91)] were increased, Vimentin protein expression level was in-creased, E-cadherin protein expression level was decreased, the difference was statistically significant (P<0.001).Conclusion Down-regulated of lncRNA SNHG7 inhibits the migration and invasion of ovarian cancer SK-OV-3 cells, and the mechanism is related to the targeted regulation of miR-34a.
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