BBC3基因表达与肺鳞状细胞癌预后的相关性及其分子机制研究

摘要

目的 验证BCL2绑定组件3(BBC3)基因表达与肺鳞状细胞癌(LUSC)术后生存时间的相关性,探究其分子机制.方法 利用实时荧光定量PCR(qRT-PCR)技术检测BBC3基因在39例LUSC患者肿瘤组织标本内的表达,并对患者进行随访,收集临床生存资料.使用Kaplan-Meier法结合log rank检验进行生存分析,使用COX比例风险模型进行多因素生存分析.在NCI-H226细胞系内过表达BBC3基因,利用噻唑蓝(MTT)法及流式细胞术检测其表达对细胞增殖及凋亡的影响.结果 BBC3的表达与肿瘤是否转移(r=0.556,P=0.023)、肿瘤大小(r=0.532,P=0.042)、T分期(r=0.551,P=0.021)及TNM分期(r=0.524,P=0.047)明显相关.Kaplan-Meier法结合log rank检验发现BBC3表达与患者生存时间明显相关,BBC3高表达者生存时间明显高于BBC3低表达者(x2=7.542,P=0.006).COX比例风险模型分析发现,肿瘤是否转移、肿瘤T分期、TNM分期及BBC3表达均独立且明显影响患者的生存时间(P＜0.05).重组BBC3质粒组凋亡细胞比例明显高于对照组与空载质粒组,差异均有统计学意义(P＜0.05).结论 BBC3基因表达可抑制肿瘤细胞增殖、促进凋亡,与LUSC预后具有明显相关性,可能作为判别LUSC术后生存预测的的潜在辅助性分子标志物.%Objective To explore the correlation between BCL2 binding component 3 (BBC3) expression and postoperative survival time of lung squamous cell carcinoma (LUSC),and investigate its molecular mechanism.Methods The expression of BBC3 in 39 patients was detected by using qRT-PCR assay.Meanwhile,clinical data of all patients were collected by follow-up visit.Then,the survival analysis was performed by using Kaplan-Meier and log rank tests.Moreover,multiple factors analysis was performed using COX proportional hazard model.At last,BBC3 was over-expressed in NCI-H226 cell lines,then detected the effects of BBC3 expression on cell proliferation and apoptosis by using MTT assay and flow cytometry.Results BBC3 expression were significantly correlated with the tumor metastasis (r=0.556,P=0.023),tumor size (r=0.532,P =0.042),T staging (r =0.551,P=0.021) and TNM staging (r=0.524,P=0.047).Meanwhile,the results of Kaplan-Meier and log rank tests found that BBC3 expression was significantly correlated with survival time of patients with LUSC,and the length of survival time in patients with high BBC3 expression was longer than that in patients with low BBC3 expression (x2 =7.542,P=0.006).The COX proportional hazard model indicated that tumor metastasis,T staging,TNM staging and BBC3 expression were independent factors which significantly affected survival time.Moreover,the proportion of apoptotic cells in the recombinant plasmid BBCs group was higher than that in the empty plasmid group and control group (P＜0.05).Conclusion BBC3 expression could suppress the proliferation of tumor cells and promote apoptosis,and are significantly correlated with survival time of patients,so which may be assistant biomarkers for prognosis of LUSC.