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下调TRIM33基因对人胃癌细胞BGC-823增殖和侵袭的影响
作者:王文莙 王菲 张弛 杨媛媛 马艳梅 徐远义 曹相玫 
单位:750004 银川 宁夏医科大学基础医学院病理学系(王文莙、王菲、张弛、杨媛媛、马艳梅、徐远义、曹相玫) 
关键词:胃癌 转染 TRIM33 增殖 侵袭 
分类号:R73
出版年,卷(期):页码:2019,44(3):210-214
摘要:

[摘要]  目的  观察三结构域家族33(TRIM33)基因下调对人胃癌细胞BGC-823增殖和侵袭能力的影响,探讨TRIM33基因在胃癌进展中的作用。方法  将人胃癌细胞BGC-823分为空载组(对照组)si-TRIM33基因转染组(si- TRIM33)。对照组以脂质体Lipofectamine 2000为载体转入空载体,si-TRIM33组用重组质粒si-TRIM33转染。采用Western blotting检测TRIM33蛋白的表达水平,CCK-8EdU法检测细胞增殖能力,平板克隆形成实验检测细胞克隆形成率,划痕实验检测细胞侵袭能力。结果  转染si-TRIM33BGC-823细胞TRIM33蛋白表达水平明显下调(P<0.01)CCK-8实验结果显示,培养2448hsi-TRIM33组细胞增殖能力(分别为0.75±0.021.16±0.10)与对照组(分别为0.48±0.060.51±0.11)比较均明显增强(P<0.01)EdU实验结果显示,si-TRIM33组细胞增殖率[(42.92±5.05)%]明显高于对照组[(14.43±1.79)%P<0.01]。平板克隆形成实验结果显示,si-TRIM33组克隆球数量[(1233.00±242.62)] 明显多于对照组[(675.00±141.52)个,P<0.05]。侵袭实验中,转染12h,对照组和si-TRIM33组细胞划痕愈合率分别为(10.5±1.8)%(16.9±3.2)%,转染24h划痕愈合率分别为(16.7±1.6)%(25.8±3.1)%si-TRIM33组划痕愈合率与对照组比较明显升高(P<0.05)结论  下调TRIM33基因可增强人胃癌细胞BGC-823的增殖和侵袭能力。

[Abstract]  Objective  To explore the potential function of TRIM33 in the progression of gastric cancer using human gastric cancer BGC-823 cells with down-regulated TRIM33. Methods  Recombinant plasmid si-TRIM33 and control vector were transfected into BC-823 cells using Liposome 2000. The protein level of TRIM33 was detected by Western blotting. CCK-8 and EdU assay were used to evaluate cell proliferation. Plate cloning assay was employed to exam the clone formation rate and the scratch test was used to assess cell invasion ability. Results  Comparing to cells that transfected with the control vector (control group), cells transfected with si-TRIM33 vectors (si-TRIM33 group) showed dramatically decreased levels of TRIM33 expression (P<0.01). Cells transfected with the control vectors showed the OD value of 0.48±0.06 and 0.51±0.11 at 24h and 48h, respectively; while cells transfected with si-TRIM33 vectors showed the OD values of 0.75±0.02 and 1.16±0.10 at 24h and 48h, respectively, using CCK-8 test. Therefore, knocking down TRIM3 in BGC-823 cells promotes cell proliferation (P<0.01). This is further confirmed using EdU assay. The cell proliferation rates of cells transfected with control vector and cells with the si-TRIM33 vectors were 14.43±1.79 and 42.92±5.05, respectively (P<0.01). In the plate cloning formation experiment, the number of spheres in the control group and the si-TRIM33 group were 675.00±141.52 and 1233.00±242.62, respectively. This showed that the down-regulation of TRIM33 enhances the colony formation rate (P<0.05). In the invasion experiment, scratch healing rates of the control group and the si-TRIM33 group at 12h were 0.105±0.018 and 0.169±0.032, respectively. The healing rates at 24h in the control group and the si-TRIM33 group were 0.167±0.016 and 0.258±0.031, respectively. Together, decreased TRIM33 enhances cell invasion (P<0.05). Conclusion  Down-regulation of TRIM33 expression promotes proliferation and invasion of human gastric cancer BGC-823 cells.

基金项目:
国家自然科学基金(81560501);宁夏回族自治区2017留学人员创新创业项目择优资助(宁人社函[2017]84号);宁夏重点研发项目(2018BEG03010)
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