Chai Yaru, Ji Penghui, Effah Clement Yaw, Duan Shuyin, Feng Feifei, Wu Yongjun
College of Public Health, Zhengzhou University, Zhengzhou 450001, Henan, China.
Henan Center for Disease Control and Prevention, Zhengzhou 450016, Henan, China.
Toxicol Res (Camb). 2023 Mar 13;12(2):270-281. doi: 10.1093/toxres/tfad015. eCollection 2023 Apr.
Coal tar pitch extract (CTPE) was carcinogenic and could cause occupational lung cancer. Hence, we explored the changes of protein molecules during CTPE-induced malignant transformation (MT) of immortalized human bronchial epithelial (BEAS-2B) cells and provided clues for screening early biomarkers of CTPE-associated occupational lung cancer. The MT model of BEAS-2B cells induced by CTPE with 15.0 μg/mL. Subsequently, the MT of the BEAS-2B cells was verified by morphological observation, cell proliferation test, plate colony formation assay, and cell cycle assay. At the end of the experiment, we explored the differentially expressed proteins (DEPs) by total protein tandem mass tags quantitative proteomics technique between DMSO40 cells and CTPE40 cells. It was found that the proliferation ability, and colony formation rate were enhanced, and the cell cycle was changed. Then, bioinformatics analysis showed that a total of 107 DEPs were screened between CTPE40 and DMSO40 cells, of which 74 were up-regulated and 33 were down-regulated. As a result, 6 hub proteins were screened by protein-protein interaction network analysis. The expression levels of COX7A2, COX7C, MT-CO2, NDUFB4, and NDUFB7 were up-regulated as well as the expression of RPS29 protein was down-regulated. In summary, we established an MT model and explored the changes in protein molecules. As a result, this study suggested that changes of protein molecules, including COX7A2, COX7C, NDUFB7, MT-CO2, NDUFB4, and RPS29, occurred at the stage of BEAS-2B cell malignancy following CTPE exposure, which provided key information for screening biomarkers for CTPE-related occupational lung cancer.
煤焦油沥青提取物(CTPE)具有致癌性,可导致职业性肺癌。因此,我们探究了永生化人支气管上皮(BEAS-2B)细胞在CTPE诱导的恶性转化(MT)过程中蛋白质分子的变化,为筛选CTPE相关职业性肺癌的早期生物标志物提供线索。用15.0μg/mL的CTPE诱导BEAS-2B细胞的MT模型。随后,通过形态学观察、细胞增殖试验、平板集落形成试验和细胞周期试验验证BEAS-2B细胞的MT。在实验结束时,我们采用全蛋白串联质量标签定量蛋白质组学技术探究了二甲基亚砜(DMSO)处理40代细胞(DMSO40细胞)和CTPE处理40代细胞(CTPE40细胞)之间的差异表达蛋白(DEP)。结果发现,细胞的增殖能力和集落形成率增强,细胞周期发生改变。然后,生物信息学分析表明,CTPE40细胞和DMSO40细胞之间共筛选出107个DEP,其中74个上调,33个下调。结果,通过蛋白质-蛋白质相互作用网络分析筛选出6个枢纽蛋白。细胞色素c氧化酶7A2(COX7A2)、细胞色素c氧化酶7C(COX7C)、线粒体细胞色素c氧化酶亚基Ⅱ(MT-CO2)、NADH脱氢酶(泛醌)铁硫蛋白4(NDUFB4)和NADH脱氢酶(泛醌)铁硫蛋白7(NDUFB7)的表达水平上调,而核糖体蛋白S29(RPS29)的表达下调。综上所述,我们建立了一个MT模型并探究了蛋白质分子的变化。结果,本研究表明,CTPE暴露后BEAS-2B细胞恶变阶段发生了包括COX7A2、COX7C、NDUFB7、MT-CO2、NDUFB4和RPS29在内的蛋白质分子变化,这为筛选CTPE相关职业性肺癌的生物标志物提供了关键信息。