长链非编码RNA-scarna 4在膀胱尿路上皮癌的作用研究

doi:10.3969/j.issn.1674-490X.2018.02.004

摘要/Abstract

摘要： 目的观察长链非编码RNA-scarna 4(LncRNA-scarna 4)在膀胱尿路上皮癌的表达,探讨抑制其表达对癌细胞的生长活力、凋亡和增殖活动的影响,明确其调控通路。方法收集尿路上皮癌组织及癌旁组织标本(n=6)、T24和HT1376尿路上皮癌细胞及正常尿路上皮细胞SV-HUC-1,qRT-PCR检测其LncRNA-scarna 4表达。分别将T24及HT1376细胞分为3组:Lnc-S组转染LncRNA-scarna 4的siRNA,Lnc-C组转染随机序列siRNA,C组与单纯培养基培养。通过台盼蓝拒染法鉴定细胞生长活力、Hoechst33342凋亡染色及Western Blot检测凋亡蛋白Bcl2表达,观察细胞凋亡活动、软琼脂集落形成实验观察肿瘤细胞增殖活动;Western Blot观察PI3K/Akt通路蛋白表达。结果 LncRNA-scarna 4在膀胱癌组织、T24及HT1376细胞表达显著增高(P<0.05)。转染抑制LncRNA-scarna 4表达导致Lnc-S组活细胞率显著降低(T24:42.1%±11.2% vs. 87.3%±8.5%;HT1376:35.3%±8.2% vs. 81.4%±10.4%,P<0.05),凋亡细胞数(/10³细胞)显著增加(T24:88±11 vs. 41±9;HT1376:79±8 vs. 47±8,P<0.05),凋亡蛋白Bcl2表达亦显著增加(T24:181.4%±20.6% vs. 119.6%±11.3%;HT1376:213.3%±34.6% vs. 121.4%±24.4%,P<0.05),克隆形成数显著降低(T24:2.6±1.1 vs. 12.5±4.6;HT1376:3.6±2.0 vs. 12.1±2.0,P<0.05),增殖及迁移PI3K/Akt通路蛋白表达显著降低(P<0.05)。结论 LncRNA-scarna 4在膀胱尿路上皮癌表达异常增高,通过抑制其表达促进癌细胞凋亡、抑制其增殖能力,其作用通路是PI3K/Akt通路。

关键词: 长链非编码RNA, 尿路上皮癌, 凋亡, 增殖

Abstract: Objective To observe the expression mode of long chain non-coding RNA-scarna 4(LncRNA-scarna 4)in bladder urothelial carcinoma and the modulation effects of LncRNA-scarna 4 in T24 and HT1376 cancer cell lines.Methods Samples were collected from bladder urothelial carcinoma tissue(n=6), para-carcinoma tissue(n=6)and T24, HT1376 and SV-HUC-1 cell lines. And the expression of LncRNA-scarna 4 was evaluated by qRT-PCR. T24 and HT1376 cells were cultured and treated with three different processing: Lnc-C group(infected with random sequence siRNA), Lnc-S group(infected with LncRNA-scarna 4 sequence siRNA), and C group(cultured with medium). To detect17 the vitality, apoptotic behavior, and proliferation of these cells, typan blue exclusion test, Hoechst33342 staining and western blot for apoptotic protein Bcl2, and soft agar colony formation assay were used respectively. Western blot method was also used to measure the expression of PI3K/Akt pathway. Results LncRNA-scarna 4 was significantly up-regulated both in bladder carcinoma and T24 and HT1376 cells(P<0.05). After infection, the viable rate was significantly lower in Lnc-S group than Lnc-C group(T24: 42.1%±11.2% vs. 87.3%±8.5%, HT1376: 35.3%±8.2% vs. 81.4%±10.4%, P<0.05). The apoptotic number(/103cells)was significantly increased in Lnc-S group(T24: 88±11 vs. 41±9; HT1376: 79±8 vs. 47±8. P<0.05), and the expression of Bcl2 protein was also increased(T24: 181.4%±20.6% vs. 119.6%±11.3%; HT1376: 213.3%±34.6% vs. 121.4%±24.4%, P<0.05). The colony number was lower in Lnc-S group(T24: 2.6±1.1 vs. 12.5±4.6; HT1376:3.6±2.0 vs. 12.1±2.0, P<0.05). Both the PI3K and Akt protein were significantly decreased in Lnc-S group(P<0.05). Conclusion LncRNA-scarna 4 is aberrantly expressed in bladder carcinoma, which participates in the process of apoptotic facilitation, abnormal proliferation through the PI3K/Akt signal pathway.

Key words: LncRNA, urothelial carcinoma, apoptosis, proliferation

中图分类号:

R394

何廉波,杨磊. 长链非编码RNA-scarna 4在膀胱尿路上皮癌的作用研究[J]. 医学研究与教育, 2018, 35(2): 26-31.

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