摘要
目的探讨组蛋白去乙酰化酶抑制剂罗米地辛对肺癌多倍体肿瘤巨细胞(PGCC)生物学行为的影响。方法人非小细胞肺癌细胞系NCI-H1299购自中国科学院上海细胞库。用100 nmol/L多西他赛处理NCI-H1299细胞24 h,建立肺癌PGCC模型,作为PGCC组;用100 nmol/L罗米地辛处理PGCC组细胞48 h,记为PGCC+罗米地辛组;将二甲基亚砜(DMSO)处理组细胞记为DMSO组。采用流式细胞术检测细胞DNA含量;细胞计数试剂盒(CCK-8)法检测细胞增殖;Transwell法检测细胞迁移能力;流式细胞术检测线粒体膜电位;Western blot检测细胞中存活素蛋白(Survivin)、骨髓细胞瘤病毒癌基因同源物蛋白(c-Myc)、波形蛋白(Vimentin)、B细胞淋巴瘤/白血病-x基因长片段蛋白(Bcl-xl)和B细胞淋巴瘤/白血病-2相关X蛋白(Bax)蛋白表达水平。结果流式细胞术检测细胞DNA含量结果显示,PGCC组PGCC亚群(DNA含量>4 N)百分比[(60.53±9.92)%]明显高于DMSO组[(2.08±0.62)%],差异有统计学意义(P<0.05)。CCK-8检测结果表明,随着罗米地辛药物浓度逐渐增加,其对细胞抑制率也逐渐增高,同时,罗米地辛处理肺癌PGCC 48 h对细胞抑制率高于24 h,差异有统计学意义(P<0.05)。Transwell检测结果显示,PGCC+罗米地辛组细胞迁移数[(13±1)个]明显少于PGCC组[(51±2)个];FACScantoⅡ流式细胞仪检测样本结果显示,PGCC+罗米地辛组的红色荧光/绿色荧光比值(2.31±0.10)明显低于PGCC组(4.55±0.52);Westernblot检测结果显示,与PGCC组相比,PGCC+罗米地辛组中Survivin、c-Myc、Vimentin和Bcl-xl表达水平明显下调,Bax表达上调,差异均有统计学意义(P<0.05)。结论罗米地辛能够抑制肺癌PGCC增殖和迁移能力,并促进肺癌PGCC凋亡。
Objective To investigate the effect of histone deacetylase inhibitor Romidepsin on biological behavior of polyploid giant cancer cell(PGCC)in lung cancer.Methods Human non-small cell lung cancer cell line NCI-H1299 was purchased from Shanghai Cell Bank of Chinese Academy of Sciences.NCI-H1299 cells were treated with 100 nmol/L Docetaxel for 24 hours,The lung cancer PGCC model was established,as PGCC group,the PGCC group cells were treated with 100 nmol/L Romidepsin for 48 h recorded as the PGCC+Romidepsin group.The cells were treated with dimethyl sulfoxide(DMSO)recorded as the DMSO group.DNA content was detected by flow cytometry.Cell proliferation was detected by cell counting kit-8(CCK-8)assay.Cell migration ability was detected by Transwell method.Mitochondrial membrane potential was detected by flow cytometry.Protein expression levels of Survivin,c-Myc,Vimentin,Bcl-xl and Bax were detected by Western blot.Results The results of DNA content detection by flow cytometry showed that the percentage of polyploid giant cancer cell subgroup(DNA content>4 N)in PGCC group[(60.53±9.92)%]was higher than that in DMSO group[(2.08±0.62)%],and the difference was statistically significant(P<0.05).The results of CCK-8 showed that with the gradual increase of the concentration of Romidepsin,its inhibition rate on cells also gradually increased,at the same time,the inhibition rate of lung cancer PGCC treated with Romidepsin for 48 hours was higher than that for 24 hours,and the difference was statistically significant(P<0.05).Transwell test results showed that the number of cell migration(13±1)in PGCC+Romidixin group was significantly lower than that in PGCC group(51±2).FACSCantoⅡflow cytometry detection results showed that the red fluorescence/green fluorescence ratio of PGCC+Romidepsin group(2.31±0.10)was significantly lower than that of PGCC group(4.55±0.52).Western blot analysis showed that compared with the PGCC group,the expression levels of survivin,c-Myc,Vimentin and Bcl-xl in PGCC+Romidixin group were significantly down-regulated,while the expression of Bax was up-regulated,the differences were statistically significant(P<0.05).Conclusions Romidepsininhibites PGCC proliferation and migration,and promotes PGCC apoptosis in lung cancer.
作者
杨紫恩
邢思宁
王丽丽
刘硕
孙凌燕
于卉影
Yang Zi′en;Xing Sining;Wang Lili;Liu Shuo;Sun Lingyan;Yu Huiying(Department of Basic Medical Laboratory,General Hospital of Northern Theater Command,Shenyang 110016,China)
出处
《中国临床实用医学》
2023年第1期12-17,共6页
China Clinical Practical Medicine
基金
辽宁省科学技术计划项目(2020JH2/10300160)。
关键词
肺癌
多倍体肿瘤巨细胞
罗米地辛
Lung cancer
Polyploid giant cancer cell
Romidepsin
作者简介
通信作者:于卉影,Email:hyingy@sina.com。
