MiR-194对恶性黑色素瘤细胞增殖和凋亡的影响及机制

发布时间：2018-02-02 21:19

本文关键词： mic R- 黑色素瘤增殖凋亡　出处：《中国现代医学杂志》2017年21期 　论文类型：期刊论文

【摘要】：目的研究mi R-194对人恶性黑色素瘤细胞的增殖和凋亡的影响及机制。方法应用实时荧光定量聚合酶链反应(q RT-PCR)分别检测mi R-194在原代正常人皮肤黑色素细胞系PIG1和5种恶性黑色素瘤细胞系(A375、SK-MEL-1、SK-MEL-2、SK-MEL-5及SK-MEL-28)中的相对表达量。利用Lipofectamine TM 2000分别将A375细胞系转染mi R-194 mimics和阴性对照质粒,从而将A375细胞系分为mi R-194模拟物组和阴性对照组;应用MTT法和流式细胞术分别测定mi R-194模拟物组和阴性对照组细胞的增殖能力及凋亡率,蛋白印迹(Western blot)分别检测mi R-194模拟物组和阴性对照组的细胞周期蛋白D1(Cyclin D1)、裂解型半胱氨酸天冬氨酸蛋白酶3(Cleaved Caspase-3)蛋白表达情况。结果 mi R-194在5种黑色素瘤细胞系A375、SKMEL-1、SK-MEL-2、SK-MEL-5及SK-MEL-28中的表达水平低于正常人皮肤黑色素细胞系PIG1,依次分别是正常细胞系的(0.18±0.03)、(0.25±0.05)、(0.37±0.03)、(0.39±0.04)及(0.45±0.03)倍(均P0.05)。转染后第0、1、2、3、4及5天,mi R-194模拟物组vs阴性对照组的O D值分别为:(0.18±0.02)vs(0.19±0.03)(P0.05)、(0.29±0.02)vs(0.27±0.03)(P0.05)、(0.42±0.08)vs(0.45±0.07)(P0.05)、(0.63±0.09)vs(1.17±0.12)(P0.05)、(1.05±0.15)vs(2.15±0.21)(P0.05)及(1.87±0.23)vs(3.18±0.27)(P0.05)。mi R-194模拟物组和阴性对照组的细胞凋亡率分别为24.2%和9.3%(P0.05)。Cyclin D1和Cleaved Caspase-3蛋白在mi R-194模拟物组中的表达量分别是阴性对照组的(0.36±0.04)和(3.2±0.28)倍(均P0.05)。结论 mi R-194在黑色素瘤细胞中低表达,mi R-194表达上调可促进黑色素瘤细胞凋亡并抑制黑色素瘤细胞的增殖,其机制可能为下调Cyclin D1蛋白及上调Cleaved Caspase-3蛋白表达。
[Abstract]:Objective to study the effect and mechanism of miR-194 on proliferation and apoptosis of human malignant melanoma cells. Mi R-194 was detected in primary normal human melanoma cell line PIG1 and 5 malignant melanoma cell lines, A375, respectively. SK-MEL-1, SK-MEL-2. SK-MEL-5 and SK-MEL-28). A375 cell line was transfected into mi R-194 mimics and negative control plasmid by Lipofectamine TM2000. A375 cell line was divided into miR-194 analogue group and negative control group. MTT assay and flow cytometry were used to determine the proliferative ability and apoptosis rate of mitR-194 analogue group and negative control group, respectively. Western blot was used to detect cyclin D1cyclin D1 in miR-194 mimetic group and negative control group. Results the expression of caspase-3 protein was detected in 5 melanoma cell lines A375. The expression levels of SK-MEL-5 and SK-MEL-28 in SKMEL-1 and SK-MEL-2 were lower than those in normal human melanoma cell line PIG1. The normal cell lines were 0.18 卤0.03, 0.25 卤0.05, 0.37 卤0.03, respectively. 0. 39 卤0. 04) and 0. 45 卤0. 03) times (P 0. 05). The O-D values of the miR-194 analogue group vs the negative control group were 0.18 卤0.02 卤0.02 vs 0.19 卤0.03 respectively (P 0.05). 0. 29 卤0. 02? VSD 0.27 卤0. 03? P0. 05??? P0. 42 卤0. 08? VSD 0.45 卤0. 07? P0. 05). 0.63 卤0.09 VSD 1.17 卤0.12 (P0.05). 1. 05 卤0. 15 卤0. 15 卤0. 21 (P0.05) and 1. 87 卤0. 23 VSD 3. 18 卤0. 27 (P0.05) (P < 0. 05, P 0. 05, P 0. 05, P 0. 05, P 0. 05). The apoptotic rates in the miR-194 analogue group and the negative control group were 24.2% and 9.3 respectively (P 0.05). The expression of Cyclin D1 and Cleaved Caspase-3 protein in the miR-194 analog group was 0.36 卤0.04) and (. Conclusion the expression of miR-194 is low in melanoma cells. The up-regulated expression of miR-194 could promote the apoptosis of melanoma cells and inhibit the proliferation of melanoma cells. The mechanism may be down-regulation of Cyclin D1 protein and up-regulation of Cleaved Caspase-3 protein expression.
【作者单位】：华中科技大学同济医学院附属武汉中心医院皮肤科;湖北省武汉市第一医院肿瘤科;
【分类号】：R739.5
【正文快照】： 恶性黑色素瘤是一类恶性程度较高的皮肤肿瘤[1-2],常见于皮肤,约占皮肤肿瘤的10%,死亡率可接近80%[3-4],其发病率在全球呈不断上升趋势[3-4],有家族聚集倾向[3-4]。本病早期症状不典型,我国每年新发病例持续增长约2万例[2],本病易于早期转移,预后多较差,5年生存率仅约15%[2]。

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