| 张炜,魏珍星,郭洁,等.微小 RNA?129?5p在喉鳞状细胞癌中的表达及对细胞增殖、侵袭的影响[J].安徽医药,2021,25(1):22-27. |
| 微小 RNA?129?5p在喉鳞状细胞癌中的表达及对细胞增殖、侵袭的影响 |
| The expression of miR?129?5p in laryngeal squamous cell carcinoma and its effects on cell proliferation and invasion |
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| DOI：10.3969/j.issn.1009?6469.2021.01.006. |
| 中文关键词: 喉肿瘤 转染 microRNA?129?5p 细胞周期蛋白 D1 人喉表皮样癌细胞(Hep?2细胞) 基质金属蛋白酶类 增殖 侵袭 |
| 英文关键词: Laryngeal neoplasms Transfection MicroRNA?129?5p Cyclin D1 Human epithelial type 2(HEp?2) Matrix metalloproteinases Proliferation Invasion |
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| 中文摘要: |
| 目的研究微小 RNA?129?5p在喉鳞状细胞癌中的表达及对细胞增殖、侵袭的影响。方法采用实时定量实时荧光定量 PCR仪(RT?qPCR)检测喉鳞状细胞癌、癌旁组织及喉鳞状细胞癌 Hep?2细胞中 miR?129?5p表达水平，转染 miR?129?5p模拟物(mimics)喉鳞状细胞癌 Hep?2细胞设为 mimics组，转染 miR?129?5p mimics空白对照组(NC)喉鳞状细胞癌 Hep?2细胞设为 mimics?NC组，将未转染喉鳞状细胞癌 Hep?2细胞设为 NC组。采用四甲基偶氮唑蓝(MTT)法、平板克隆试验检测各组喉鳞状细胞癌 Hep?2细胞增殖情况；采用 Transwell各组喉鳞状细胞癌 Hep?2细胞侵袭情况；采用蛋白质印迹法试验检测各组喉鳞状细胞癌 Hep?2细胞增殖、侵袭相关蛋白表达水平。结果与癌旁组织(1.03±0.14)比较，喉鳞状细胞癌组织中 miR?129?5p(0.48±0.08)表达水平显著降低(P＜0.05)。同一时间和不同时间 NC组、 mimic?NC组miR?129?5p表达水平、喉鳞状细胞癌 Hep?2细胞增殖抑制率、平板克隆形成率、侵袭细胞数、一抗鼠源原癌基因(c?Myc)、细胞周期蛋白 D1(Cyclin D1)、基质金属蛋白酶 2(MMP?2)、基质金属蛋白酶 9(MMP?9)蛋白水平比较差异无统计学意义(P＞0.05)。同一时间与 NC组、 mimics?NC组比较， mimics组miR?129?5p表达水平、喉鳞状细胞癌 Hep?2细胞增殖抑制显著增加(P＜0.05)，与 24 h比较， 48 h mimics组 miR?129?5p表达水平、喉鳞状细胞癌 Hep?2细胞增殖抑制显著增加(P＜0.05)。平板克隆形成率为(98.96±10.60)%、(97.57±10.82)%、(46.91±9.03)%、侵袭细胞数为(235.62±36.18)、(227.31±35.90)、(94.38±19.86)个、 c?Myc(3.92±0.71)、(3.89±0.60)、(2.18± 0.46)、 Cyclin D1(1.54±0.27)、(1.52±0.30)、(0.39±0.08)、 MMP?2(1.12±0.28)、(1.11±0.25)、(0.36±0.07)、 MMP?9(1.03±0.14)、(0.98±0.13)、(0.29±0.05)蛋白水平均显著降低(均 P＜0.05)。结论 miR?129?5p在喉鳞状细胞癌组织中低表达，上调 miR?129?5p 可抑制喉鳞状细胞癌 Hep?2细胞增殖及侵袭。 |
| 英文摘要: |
| Objective To study the expression of miR?129?5p in laryngeal squamous cell carcinoma and its effects on cell prolif? eration and invasion.Methods Real?time quantitative PCR(RT?qPCR)was used to detect the expression of miR?129?5p in laryn? geal squamous cell carcinoma，para?cancerous tissues and Hep?2 cells of laryngeal squamous cell carcinoma.MiR?129?5p mimicstransfected laryngeal squamous cell carcinoma Hep?2 cells were assigned to mimics group，Hep?2 cells of laryngeal squamous cellcarcinoma transfected with miR?129?5p mimics?NC blank control group were set as mimics?NC group，and the untransfected laryn?geal squamous cell carcinoma Hep?2 cells were assigned to NC group.The proliferation of laryngeal squamous cell carcinoma Hep?2cells was detected by MTT assay and plate cloning test，the invasion of Hep?2 cells in each group of laryngeal squamous cell carci? noma was studied by Transwell，Western blot(WB)was used to detect the expressions of proliferation and invasion?related pro?teins in Hep?2 cells of laryngeal squamous cell carcinoma.Results Compared with the para?cancerous tissues(1.03±0.14)，the ex? pression level of miR?129?5p(0.48±0.08)in laryngeal squamous cell carcinoma was significantly lower(P＜0.05).There were no significant differences in the expression of miR?129?5p，the inhibition rate of proliferation of laryngeal squamous cell carcinoma Hep?2 cells，the rate of plate cloning，the number of invasive cells，c?Myc，Cyclin D1，matrix metalloproteinase 2(MMP?2)and matrix metalloproteinase 9(MMP?9)protein levels between NC group and mimic?NC group at the same time and at different time(P＞0.05).At the same time，compared with NC group and mimics?NC group，the expression of miR?129?5p and the proliferation inhibi?tion of laryngeal squamous cell carcinoma Hep?2 cells in mimics group increased significantly(P＜0.05).Compared with 24h，the expression of miR?129?5p and the proliferation inhibition of laryngeal squamous cell carcinoma Hep?2 cells in 48h mimics group in?creased significantly(P＜0.05).Plate cloning rate(98.96±10.60)，(97.57±10.82)，(46.91±9.03)%，number of invasive cells(235.62±36.18)(227.31±35.90)(94.38±19.86)individual，c?Myc(3.92±0.71)(3.89±0.60)(2.18±0.46)Cyclin D1(1.54±0.27)(1.52±0.30(0.39±0.08)MP?2(1.12±0.28)(1.11±0.25)，MMP?2(1.12±0.28)，(1.11±0.25)，(0.36±0.07)and MMP?9(1.03±0.14)，(0.98±0.13)，(0.29±0.05)protein levels were significantly decreased(P＜0.05).Conclusions The expressionofmiR?129?5pislow，mous cell carcinoma tissues.Up?regulation of miR?129?5p can inhibit the proliferation and invasion of laryngeal squamous cell carci?noma Hep?2 cells. |
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