长链非编码RNA HOTAIRM1对MPP+处理后MN9D细胞活力和凋亡的影响

doi:10.3969/j.issn.1004-616x.2025.02.006

摘要/Abstract

摘要： 目的：探讨长链非编码RNA(lncRNA)骨髓特异性HOX基因转录本反义RNA 1(HOTAIRM1)对1-甲基-4-苯基吡啶离子(MPP⁺)处理后的MN9D细胞活力和凋亡的影响。方法：将不同浓度(0、0.25、0.5、1、1.25、2.5 mmol/L)的MPP⁺作用于MN9D细胞不同时间(0、24、48和72 h)后，采用CCK-8检测各组细胞活力，选择MPP⁺最适浓度和最佳作用时间进行后续试验。选用siRNA进行转染，将MN9D细胞分为对照组、MPP⁺处理组(1 mmol/L MPP⁺处理24 h)、siRNA对照组(siRNA阴性对照序列转染1 h后用 1 mmol/L MPP⁺处理24 h)，和siR-HOTAIRM1干扰组(siR-HOTAIRM1转染1 h后用1 mmol/L MPP⁺处理24 h)。分别采用CCK-8检测各组细胞活力；实时荧光定量逆转录PCR(qRT-PCR)检测各组细胞中lncRNA HOTAIRM1的表达；流式细胞术检测各组细胞凋亡情况。结果：与对照组比较，不同浓度MPP⁺处理24 h，细胞活力随浓度的升高而降低(P<0.05)，而lncRNA HOTAIRM1表达水平逐渐升高(P<0.05)；1 mmol/L MPP⁺处理不同时间后，细胞活力明显下降(P<0.05)，lncRNA HOTAIRM1表达水平先升高后降低(P<0.05)，故选择1 mmol/L MPP⁺处理24 h进行后续试验。与转染siRNA阴性对照序列组相比，siR-HOTAIRM1干扰之后，lncRNA HOTAIRM1表达显著降低(P<0.01)，但显著增强MPP⁺处理后的MN9D细胞的活力(P<0.01)；明显抑制MPP⁺处理后的MN9D细胞的凋亡(P<0.01)，从而保护MN9D细胞。结论：干扰lncRNA HOTAIRM1的表达可增强MPP⁺处理后的MN9D细胞活力，并抑制细胞凋亡。lncRNA HOTAIRM1敲低对MPP⁺处理后的MN9D细胞损伤具有保护作用。

关键词: MN9D细胞, 帕金森病, 长链非编码RNA, HOTAIRM1, 细胞凋亡

Abstract: OBJECTIVE：To explore the impacts of the long non-coding RNA (lncRNA) HOX transcript antisense RNA，myeloid-specific 1 (HOTAIRM1)，on the vitality and apoptosis of MN9D cells which were induced by 1-methyl-4-phenylpyridinium ion ([MPP⁺]). METHODS：MN9D cells were treated with different concentrations (0，0.25，0.5，1，1.25，2.5 mmol/L) of MPP⁺ for various time periods (0，24，48，and 72 h). Subsequently，cell viability of each group was evaluated using the CCK-8 assay. Based on the results，the optimal concentration of MPP⁺ and the most appropriate treatment duration were determined for subsequent experiments. siRNA transfections were conducted. The MN9D cells were then categorized into the control group，MPP⁺ group，si-NC+MPP⁺ group，and si-HOTAIRM1+MPP⁺ group. Cell viability of each group was measured by CCK-8 assay. Expression of lncRNA HOTAIRM1 was detected through real-time fluorescent quantitative reverse transcription-polymerase chain reaction (qRT-PCR). Apoptosis status was analyzed by flow cytometry. RESULTS：In comparison with the control group，lncRNA HOTAIRM1 was highly expressed in MN9D cells induced by MPP⁺ (P<0.05). When treated with 1 mmol/L MPP⁺ for 24 h，the cell viability was significantly decreased，while the expression level of lncRNA HOTAIRM1 was remarkably elevated. Hence，1 mmol/L MPP⁺ for 24 h treatment was selected for subsequent experiments. After the induction and siRNA interference，expression of lncRNA HOTAIRM1 was significantly reduced (P<0.001)，and viability of the cells were significantly enhanced (P<0.01). Knockdown of lncRNA HOTAIRM1 inhibited the apoptosis of the induced cells (P<0.01). CONCLUSION：Interfering with the expression of lncRNA HOTAIRM1 augmented viability of the MN9D cells induced by MPP⁺ and suppressed cell apoptosis. Knockdown of lncRNA HOTAIRM1 exerted a protective effect on the induced cells.

Key words: MN9D cell, Parkinson's disease, long non-coding RNA, HOTAIRM1, apoptosis

中图分类号:

R741

杨敏丽, 张艺, 高含, 谭启涛, 郑展越, 孙天奥, 潘明炼, 马勇杰, 孙艳. 长链非编码RNA HOTAIRM1对MPP⁺处理后MN9D细胞活力和凋亡的影响[J]. 癌变·畸变·突变, 2025, 37(2): 128-133.

YANG Minli, ZHANG Yi, GAO Han, TAN Qitao, ZHENG Zhanyue, SUN Tianao, PAN Minglian, MA Yongjie, SUN Yan. Expression of the long non-coding RNA HOTAIRM1 on viability and apoptosis of MPP⁺-induced MN9D cells[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2025, 37(2): 128-133.

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