| 黄荣,樊明湖,黄芬,等.长链非编码 RNA膀胱癌相关转录物 1对微小 RNA-503-5p的靶向关系及对结直肠癌细胞增殖和凋亡的影响[J].安徽医药,2021,25(8):1637-1642. |
| 长链非编码 RNA膀胱癌相关转录物 1对微小 RNA-503-5p的靶向关系及对结直肠癌细胞增殖和凋亡的影响 |
| Relationship of long-chain non-coding RNA BLACAT1 to microRNA -503-5p and the effect of colorectal cancer cell proliferation and apoptosis |
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| DOI：10.3969/j.issn.1009-6469.2021.08.038 |
| 中文关键词: 结直肠肿瘤 lncRNA BLACAT1 微小 RNA-503-5p 增殖 凋亡 |
| 英文关键词: Colorectal neoplasms LncRNA BLACAT1 MiR-503-5p Proliferation Apoptosis |
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| 中文摘要: |
| 目的研究长链非编码 RNA(lncRNA)膀胱癌相关转录物 1(BLACAT1)对微小 RNA(miR)-503-5p的靶向关系及结直肠癌细胞增殖和凋亡的影响。方法实时荧光定量 PCR(qRT-PCR)检测结肠癌细胞株 SW620，LOVO，HT29和人正常结肠黏膜上皮细胞株 NCM460中 BLACAT1和 miR-503-5p的表达。在 HT29细胞中转染 si-BLACAT1、pcDNA-BLACAT1或 miR-503-5p，噻唑(MTT)检测细胞增殖，流式细胞术检测细胞凋亡，蛋白质印迹法( Western blotting)检测细胞核相关抗原 Ki-67(Ki-67)、细胞周期蓝蛋白 D1(Cyclin D1)、活化的多聚 ADP-核糖聚合酶( Cleaved PARP)和活化的含半胱氨酸的天冬氨酸蛋白水解酶 -3(Cleaved caspase-3)的表达， starbase预测和双荧光素酶报告分析 BLACAT1与 miR-503-5p之间的靶向关系。 si-BLACAT1和 anti-miR503-5p共转染，观察干扰 miR-503-5p对沉默 BLACAT1诱导的结直肠癌细胞增殖和凋亡的影响。结果与 NCM460细胞比较， SW620、LOVO和 HT29中 BLACAT1表达量明显增加［(4.93±0.58)、(5.66±0.53)、(6.17±0.66)比(1.03±0.22)］miR-503-5p表达量减少［( 0.72±0.11)、(0.67±0.09)、(0.51±0.08)比( 1.04±0.14)］(P<0.05)。沉默 BLACAT1或转染 miR-503-5p明显，减少 HT29细胞的细胞存活率、 Ki-67、CyclinD1蛋白表达量，提高细胞凋亡率、 Cleaved PARP和 Cleaved caspase-3蛋白水平( P<0.05)过表达 BLACAT1则反之。 BLACAT1靶向 miR-503-5p调控其表达。干扰 miR-503-5p部分逆转沉默 BLACAT1抑制结直肠癌细胞，增殖、 Ki-67、CyclinD1蛋白表达和诱导结直肠癌细胞凋亡、 Cleaved PARP、Cleaved caspase-3蛋白表达的作用。结论 lncRNA BLACAT1在表达上调，沉默 BLACAT1可通过靶向调控 miR-503-5p的表达，来抑制结直肠癌细胞增殖，并诱导细胞凋亡。 |
| 英文摘要: |
| Objective To study the targeting relationship of long-chain non-coding RNA (lncRNA) BLACAT1 to microRNA (miR) 503-5p and its effect on colorectal cancer cell proliferation and apoptosis.Methods The expressions of BLACAT1 and miR-503-5p in colon cancer cell lines SW620, LOVO, HT29 and human normal colon mucosal epithelial cell line NCM460 were detected by real-time fluorescent quantitative PCR (qRT-PCR). HT29 cells were transfected with si-BLACAT1, pcDNA-BLACAT1 or miR-503-5p. MTT wasused to detect cell proliferation, flow cytometry was used to measure apoptosis, and Western blot was used to determine expression ofnuclear-related antigen Ki-67 (Ki-67), cyclin D1 (Cyclin D1), cleaved Poly ADP-ribose polymerase (Cleaved PARP), and cleaved cysteinyl aspartate specific proteinase-3 (Cleaved caspase-3). The starbase Prediction and dual luciferase reports were used to analyze the targeting relationship between BLACAT1 and miR-503-5p. si-BLACAT1 and anti-miR-503-5p were co-transfected to observe the effect of anti-miR-503-5p interfering on the proliferation and apoptosis of colorectal cancer cells induced by silenced BLACAT1.Results Compared with NCM460 cells, the expression of BLACAT1 in SW620, LOVO and HT29 increased [(4.93±0.58), (5.66±0.53), (6.17±0.66) vs. (1.03±0.22)], and the expression of miR-503-5p evidently decreased [(0.72±0.11), (0.67±0.09), (0.51±0.08) vs. (1.04±0.14)](P< 0.05). Silencing BLACAT1 or transfecting miR-503-5p greatly reduced the cell survival rate, Ki-67, and CyclinD1 protein expression in HT29 cells, and obviously increased the apoptosis rate, Cleaved PARP and Cleaved caspase-3 protein levels (P<0.05), which were the opposite of overexpression of BLACAT1. BLACAT1 targeted miR-503-5p to regulate its expression. Interfering with miR-503-5p partially reversed the effects of silencing BLACAT1 on inhibiting the colorectal cancer cell proliferation, Ki-67, CyclinD1 protein expression and inducing colorectal cancer cell apoptosis, Cleaved PARP, and Cleaved caspase-3 protein expression.Conclusion The expression of lncRNA BLACAT1 is up-regulated. Silencing BLACAT1 can inhibit the proliferation of colorectal cancer cells and induce apoptosis by targeting the regulation of miR-503-5p expression. |
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