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背景与目的印记基因dlk1因在多种肿瘤组织中出现异常表达而受到研究者越来越多的关注,但dlk1基因与肺癌的关系尚无报道。本研究首先在肺癌组织中检测了dlk1基因的表达,并进一步利用肺癌细胞系H520对dlk1基因促进细胞增殖的分子机制进行了初步研究。方法首先,采用RT-PCR在30对非小细胞肺癌肿瘤及其配对癌旁组织中检测dlk1基因的表达。然后,克隆人源dlk1基因,转染并筛选出稳定表达dlk1基因的肺癌细胞。最后,利用CCK8法研究dlk1基因对细胞增殖能力的影响,并用Western blot技术分析细胞周期蛋白Cyclin B1的表达。结果 RT-PCR结果显示,dlk1基因在36.7%的非小细胞肺癌肿瘤组织中表达水平高于癌旁肺组织。在成功获得了稳定表达外源性dlk1基因的肺癌细胞H520-dlk1的基础上,CCK8实验及平板集落实验显示,稳定转染dlk1可以明显促进肺鳞癌细胞H520的增殖能力(P<0.05)。同时,稳定表达DLK1蛋白可以上调细胞周期蛋白Cyclin B1的表达水平(P<0.05)。结论 dlk1在非小细胞肺癌中存在异常高表达,它可以通过上调细胞周期蛋白Cyclin B1的表达,促进肺鳞癌细胞H520的增殖。提示dlk1基因的异常表达可能在肺癌的发生演进中发挥作用。
BACKGROUND & OBJECTIVE: dlk1, a marker gene, has attracted more and more attention due to its abnormal expression in various tumor tissues. However, the relationship between dlk1 gene and lung cancer has not been reported yet. In this study, the expression of dlk1 gene was detected in lung cancer tissues and the molecular mechanism of dlk1 gene proliferation was further studied by lung cancer cell line H520. Methods Firstly, the expression of dlk1 gene was detected by RT-PCR in 30 pairs of non-small cell lung cancer and their matched paracancerous tissues. Then, the human dlk1 gene was cloned and transfected into lung cancer cells stably expressing the dlk1 gene. Finally, the effects of dlk1 gene on cell proliferation were investigated by CCK8 assay, and the expression of cyclin B1 was analyzed by Western blot. Results The RT-PCR results showed that dlk1 gene was expressed in 36.7% of non-small cell lung cancer tissues compared with adjacent lung tissues. On the basis of successfully obtaining H520-dlk1 lung cancer cell line stably expressing exogenous dlk1 gene, CCK8 assay and platelet colony assay showed that stable transfection of dlk1 can significantly promote the proliferation of lung squamous carcinoma cell line H520 (P <0.05). Meanwhile, stable expression of DLK1 protein could up-regulate the expression of cyclin B1 (P <0.05). Conclusions Dlk1 is abnormally expressed in non-small cell lung cancer. It can up-regulate the expression of cyclin B1 and promote the proliferation of lung squamous carcinoma cell line H520. Suggest that the abnormal expression of dlk1 gene may play an important role in the development of lung cancer.