长链非编码RNA POT1-AS1在宫颈癌顺铂耐药中的作用研究

王程; 朱逸飞; 郑世康; 张晓然; 潘夏至; 刘明波; 刘爱军

doi:10.12435/j.issn.2095-5227.2024.004

长链非编码RNA POT1-AS1在宫颈癌顺铂耐药中的作用研究

Role of long non-coding RNA POT1-AS1 in cisplatin resistance in cervical cancer

摘要

摘要:
背景同步放化疗是晚期宫颈癌(cervical cancer，CC)的治疗方法，而顺铂(cisplatin，CDDP)是目前临床化疗的主要药物之一，但CDDP耐药的发生严重制约了其临床应用，因此针对CDDP耐药发生机制的研究对CC的治疗至关重要。
目的探索长链非编码RNA(long non-coding RNA，lncRNA)POT1-AS1在宫颈癌 CDDP耐药中的作用。
方法基于TCGA数据库，分析POT1-AS1在CC组织及正常组织中的表达情况，并分析POT1-AS1的表达量与预后的关系。采用CCK-8实验评估CDDP处理后的耐药细胞系与亲本细胞系、siPOT1-AS1和siNC组的活力并测定相应的半抑制浓度(the half maximal inhibitory concentration，IC50)。实时荧光定量PCR(Quantitative Real-time PCR，qPCR)检测POT1-AS1在耐药细胞系及相应亲本细胞系中的表达以及POT1-AS1表达与CDDP作用时间的相关性。克隆形成实验检测POT1-AS1对CDDP耐药细胞增殖情况的影响。PI/DAPI双染荧光用于评估POT1-AS1与CDDP诱导的细胞死亡间的相关性。
结果 POT1-AS1在CC肿瘤组织中高表达，并与不良预后相关。耐药细胞系的细胞活力及IC50值显著高于亲本细胞系(P＜0.001)，POT1-AS1沉默组的细胞活力及IC50值显著低于阴性对照组(negative control，NC)(P＜0.001)，差异均有统计学意义。POT1-AS1在耐药细胞系中的表达显著高于亲本细胞系(P＜0.001)，且随着CDDP给药时间的延长而增加。沉默POT1-AS1后可明显抑制耐药细胞的增殖能力(P＜0.01)。与NC组相比，POT1-AS1沉默组的CC耐药细胞死亡率明显增加(P＜0.01)，尤其是在加入CDDP作用后(P＜0.01)。
结论 POT1-AS1与宫颈癌 CDDP耐药密切相关，并通过抑制CDDP诱导的细胞死亡发挥促癌作用。

Abstract:
Background Concurrent chemoradiotherapy is the treatment for advanced cervical cancer(CC), and cisplatin (CDDP) is currently one of the main drugs in clinical chemotherapy. However, the development of CDDP resistance severely limits its clinical application, making it essential to investigate the mechanisms underlying CDDP resistance for the treatment of CC.
Objective To explore the role of POT1-AS1, a long non-coding RNA, in CDDP resistance in CC.
Methods Based on the TCGA database, we investigated the expression of POT1-AS1 in CC tissues and normal tissues, and analyzed the relationship between POT1-AS1 expression levels and prognosis. CCK-8 assay was used to compare the viability of resistant cell lines after CDDP treatment with that of parental cell lines and siPOT1-AS1 and siNC groups, and to determine the corresponding half maximal inhibitory concentration (IC50). QPCR was used to examine the expression of POT1-AS1 in resistant cell lines and corresponding parental cell lines as well as the correlation between POT1-AS1 expression and the duration time of CDDP treatment. Colony formation assays was used to examine the effect of POT1-AS1 on the proliferation of CDDP-resistant cells. PI/DAPI double staining fluorescence was used to assess the correlation between POT1-AS1 and CDDP induced cell death.
Results POT1-AS1 was highly expressed in CC tumor tissues and associated with poor prognosis. The cell viability and IC50 values of the resistant cell lines were significantly increased than those of the parental cell lines (P＜0.001), and they were significantly lower in the POT1-AS1 silenced group than in the negative control group (P＜0.001), with statistically significant differences. The expression of POT1-AS1 in resistant cell lines was significantly elevated than that in parental cell lines (P＜0.001), and it increased with prolonged CDDP administration. POT1-AS1 silencing could significantly inhibit the proliferation of drug-resistant cells (P < 0.01). The resistant cell death rate of CC was significantly increased in the POT1-AS1 silenced group compared with the negative control group (P＜0.01), especially after CDDP treatment (P＜0.01).
Conclusion POT1-AS1 is closely associated with CC CDDP resistance and plays a cancer promoting role by inhibiting CDDP mediated cell death.

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