耐辐射肺腺癌细胞模型的放射生物学鉴定OA北大核心CSTPCD
Radiobiological identification of radioresistant lung adenocarcinoma cell model
目的:体外构建耐辐射的肺腺癌细胞模型,研究与肿瘤细胞辐射抗性相关的生物学变化.方法:通过分次照射建立了耐辐射的肺腺癌细胞模型A549RR;将A549和A549RR细胞各随机分为两组,分别给予0和8 Gy辐照,检测亲本细胞与耐辐射细胞的增殖能力和对放疗的敏感性;在A549、A549RR和辐射抗性部分消失的A549RR'三个细胞系中分别设置顺铂治疗组、顺铂联合放射治疗组和空白对照组,检测细胞对化疗及联合放化疗的敏感性;同时利用流式细胞术分别检测A549和A549RR细胞在接受0和8 Gy辐照后的细胞周期变化和凋亡情况,使用彗星试验检测DNA损伤情况,检测活性氧(ROS)和8-羟基脱氧鸟苷(8-OHdG)水平,并对细胞进行衰老相关β-半乳糖苷酶染色.结果:与亲本A549细胞相比,A549RR细胞的辐射抗性显著增强(P<0.01),增殖能力显著下降(P<0.01),化疗前后细胞活力无显著差异.相比亲本A549细胞,处于G2/M期的A549RR细胞显著减少(P<0.01),处于S期的A549RR细胞显著增多(P<0.01);高剂量辐照后,亲本A549细胞发生S期阻滞,而A549RR细胞的细胞周期在辐照前后无显著差异.此外,A549RR细胞在接受辐照后相比A549细胞DNA损伤减少(P<0.01),ROS水平降低(P<0.01),辐照前后A549RR细胞中8-OHdG水平和凋亡率无显著差异;A549RR细胞中处于衰老的细胞多于亲本A549细胞(P<0.01).结论:本研究通过构建耐辐射细胞模型,探究了放疗对耐辐射肺癌细胞存活的影响及其在细胞周期、DNA损伤、凋亡、衰老以及氧化应激水平方面的变化,并有助于进一步研究肺癌放疗抵抗机制.
AIM:To establish a radioresistant lung adenocarcinoma cell model in vitro and investigate the bio-logical changes related to radiation resistance in tumor cells.METHODS:A radioresistant lung adenocarcinoma cell model namely A549RR,was established by fractionated irradiation.The A549 and A549RR cells were randomly divided into two groups,which were irradiated with 0 and 8 Gy.The proliferation ability of parental cells and radioresistant cells were detected to assess their sensitivity to radiotherapy.The A549,A549RR and A549RR'(partially radioresistant)cell lines were divided into cisplatin chemotherapy group,cisplatin chemotherapy combined with radiotherapy group and blank control group to evaluate the sensitivity to chemotherapy and radiochemotherapy.Flow cytometry was used to detect the changes in cell cycle and apoptosis after irradiation,while the comet assay was used to evaluate DNA damage.The levels of reactive oxygen species(ROS)and 8-hydroxydeoxyguanosine(8-OHdG)levels were measured,and senescence-associ-ated β-galactosidase staining was performed.RESULTS:Compared with parental A549 cells,A549RR cells exhibited enhanced radiation resistance(P<0.01),decreased proliferation capacity(P<0.01),and no significant difference in cell viability before and after chemotherapy.In comparison to A549 cells,the proportion of A549RR cells at G2/M phase was significantly decreased(P<0.01),and that at S phase was significantly increased(P<0.01).After exposed to high-dose irradiation,A549 cells displayed S phase arrest,whereas A549RR cells showed no significant difference in cell cycle be-fore and after irradiation.Additionally,A549RR cells exhibited reduced DNA damage(P<0.01)and lower levels of ROS(P<0.01)compared with A549 cells following irradiation.There were no significant differences in the levels of 8-OHdG and apoptotic rate before and after irradiation in A549RR cells.CONCLUSION:In this study,by establishing a radia-tion-resistant cell model,the impact of radiotherapy on the survival of radioresistant lung cancer cells was investigated,as well as the corresponding changes in cell cycle progression,DNA damage,apoptosis,senescence,and oxidative stress levels.These findings are expected to contribute to a better understanding of the mechanisms underlying radiation resis-tance in lung cancer and may inform further research in this field.
陈家靖;刘晏娜;曾琴;金鑫;杨满意;廖明媚;赵劲风
中南大学湘雅医院卫健委纳米生物技术重点实验室,湖南 长沙 410008||河南中医药大学第二附属医院肿瘤科,河南 郑州 450046中南大学湘雅医院卫健委纳米生物技术重点实验室,湖南 长沙 410008
临床医学
肺癌辐射抗性DNA损伤细胞周期细胞凋亡细胞衰老
lung cancerradioresistanceDNA damagecell cycleapoptosiscell senescence
《中国病理生理杂志》 2024 (004)
586-593 / 8
湖南省自然科学基金资助项目(No.2022JJ30974;No.2022JJ30932)
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