| 忽平,王丽媛.长链非编码 RNA LINC00346调控微小 RNA-138-5p对宫颈癌细胞增殖、凋亡的影响[J].安徽医药,2022,26(6):1206-1210. |
| 长链非编码 RNA LINC00346调控微小 RNA-138-5p对宫颈癌细胞增殖、凋亡的影响 |
| Long non-coding RNA LINC00346 regulates the proliferation and apoptosis of cervical cancer by targeting miR-138-5p |
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| DOI：10.3969/j.issn.1009-6469.2022.06.035 |
| 中文关键词: 宫颈肿瘤 LINC00346 微小 RNA-138-5p 增殖 凋亡 |
| 英文关键词: Uterine cervical neoplasms LINC00346 MiR-138-5p Proliferation Apoptosis. |
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| 中文摘要: |
| 目的研究长链非编码 RNA(lncRNA)LINC00346影响宫颈癌细胞增殖和凋亡的分子机制。方法选取 2018年 1—12月于南阳市中心医院手术治疗的 28例宫颈癌病人的宫颈癌组织(实验组)及癌旁正常组织(对照组)为研究对象。根据细胞转染情况分为 si-NC组、 si-LINC00346组、 miR-NC组、 miR-138-5p组、 si-LINC00346+anti-miR-NC及 si-LINC00346+anti-miR-138-5p组；利用实时荧光定量逆转录聚合酶链反应( qRT-PCR)检测宫颈癌组织和宫颈癌海拉细胞中 LINC00346和微小 RNA(miR)138-5p的表达水平，蛋白质印迹法检测海拉细胞中周期素依赖激酶抑制剂 p21(P21)和胱天蛋白酶 -3(caspase-3)的蛋白含量， MTT法和流式细胞术测定细胞存活率和凋亡率，双荧光素酶报告系统验证 LINC00346和 miR-138-5p的靶向关系。结果与正常癌旁组织相比，宫颈癌组织中 LINC00346含量显著升高［( 2.51±0.08)比( 0.99±0.05)］(P<0.001)；转染 si-LINC00346(LINC00346干扰物)后，与 si-NC(阴性对照组)相比， si-LINC00346组宫颈癌海拉细胞中的 LINC00346含量显著下降［( 0.33± 0.03)比( 1.00±0.05)］P21和 caspase-3蛋白含量升高［(0.63±0.04)比( 0.22±0.02)；(0.79±0.05)比( 0.30±0.03)］，海拉细胞存活率降低［(52.84±4.39)比(，100.00±6.13)］，凋亡率升高［( 23.39±1.64)比( 8.19±1.00)］，均差异有统计学意义( P<0.001)，敲除 LINC00346可降低细胞存活率，提高细胞凋亡率及 P21和 caspase-3蛋白含量；转染野生型 WT-LINC00346的海拉细胞，与 miR-NC对照组相比， miR-138-5p组野生型 WT-LINC00346的萤火虫荧光素酶相对活性显著下降［( 0.24±0.03)比( 0.97±0.05)］。敲除 LINC00346可显著上调 miR-138-5p含量［(2.43±0.07)比( 1.01±0.05)］(P<0.001)。结论 LINC00346通过靶向 miR-138-5p调控宫颈癌海拉细胞增殖和凋亡。 |
| 英文摘要: |
| Objective To study the molecular mechanism of long non-coding RNA (lncRNA) LINC00346 affecting the proliferation and apoptosis of cervical cancer cells.Methods Cervical cancer tissues (experimental group) and normal paracancer tissues (controlgroup) of 28 patients with cervical cancer who underwent surgical treatment in Nanyang Central Hospital from January 2018 to December 2018 were selected as the research objects. According to the transfection conditions, the cells were divided into si-NC group, siLINC00346 group, miR-NC group, miR-138-5p group, si-LINC00346+anti-miR-NC group and si-LINC00346+anti-miR-138-5p group. The levels of cyclin-dependent kinase inhibitor 1A (P21) and caspase-3 proteins in HeLa cells were measured by Western blotting. Tetramethylazolium salt (MTT) assay and flow cytometry were used to determine the cell survival rate and apoptosis rate. Dual luciferasereporter assay system was performed to examine the targeting relationship between LINC00346 and miR-138-5p.Results Comparedwith that of the normal adjacent tissue, the content of LINC00346 in cervical cancer tissue group was significantly increased [(2.51±0.08) vs. (0.99±0.05)] (P<0.001). Compared with the negative control group, the LINC00346 content in cervical cancer HeLa cells in the si-LINC00346 group was significantly decreased [(0.33±0.03) vs. (1.00±0.05)], and P21 and caspase-3 protein content were significantly increased [(0.63±0.04) vs. (0.22±0.02); (0.79±0.05) vs. (0.30±0.03)], cell survival rate was decreased [(52.84±4.39) vs. (100.00± 6.13)] and apoptosis rate was increased [(23.39±1.64) vs. (8.19±1.00)], and the difference had statistical significance (P<0.001). Knockout of LINC00346 reduced the cell survival rate, increased the cell apoptosis rate and the levels of P21 and caspase-3 protein content; in HeLa cells transfected with wild-type WT-LINC00346, the relative activity of firefly luciferase in wild-type WT-LINC00346 in the miR-138-5p group was significantly decreased compared with the miR-NC control group [(0.24±0.03) vs. (0.97±0.05)]. Knockdown of LINC00346 significantly upregulated miR-138-5p content [(2.43±0.07) vs. (1.01±0.05)] (P<0.001).Conclusion LINC00346 regulates the proliferation and apoptosis of cervical cancer HeLa cells by targeting miR-138-5p. |
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