利用集落形成实验和成瘤实验初步研究新基因的成瘤作用

Primary Function Analysis of Novel Genes Using Colony Formation and Tumorigenicity Experiments

目的 探讨新基因尤其是肝癌相关新基因的功能。方法 利用集落形成实验和成瘤实验初步分析HC90和HC3898基因的功能。结果 ①HC90基因的转染能增加SMMC 772 1肝癌细胞形成的集落数量 ;②注射转染了HC90基因的SMMC 772 1细胞形成的肿瘤平均瘤重大于注射SMMC 772 1细胞的自身对照组 ;③HC3898全长cDNA转染SMMC 772 1细胞能显著抑制集落的形成 ;④注射转染HC3898基因的SMMC 772 1细胞组平均瘤重小于注射SMMC 772 1细胞的自身对照组的瘤重 ,抑瘤率为 5 0 % ;⑤HC3898实验组瘤块切片检查可见凋亡细胞及凋亡小体 ,瘤块的组织DNA经 1.5 %琼脂糖电泳分析 ,可发现典型的凋亡“阶梯状DNA条带”。结论 HC90可能是与肝癌恶性表型相关的新基因 ,转染SMMC 772 1肝癌细胞后能促进肝癌细胞的生长。HC3898基因可能诱导凋亡并抑制肝癌细胞的生长。集落形成实验和成瘤实验是对基因功能进行初步分析的有效方法 。

Purpose: To introduce a method for primary function analysis of novel genes which related to hepatocellular carcinoma(HCC). Methods: Colony formation and tumorigenicity experiments were used for primary function analysis of two HCC-related novel genes-HC90 and HC3898 cloned by our laboratory. Results: The transfection of the HC90 into the hepatocellular carcinoma cell SMMC-7721 did promote the colony formation capability. The weight of the tumor tissue derived from SMMC-7721 cells transfected with HC90 was higher than that of controls(P < 0.05). The transfection of HC3898 significantly supressed the colony formation capability of transfected SMMC-7721 cells. The tumorigenicity of HC3898 transfected SMMC-7721 hepatoma cells was reduced to 50 % as compared with vector transfected control. The DNA of the tumor tissue derived from HC3898 transfected SMMC-7721 cells displayed the typical ladder of apoptosis in electrophoresis analysis. The positive apoptotic cells were also found in tumor tissue derived from SMMC-7721 cells transfected HC3898. Conclusions: HC90 might be a growth-stimulating gene possibly related to the malignant phenotype of HCC. HC3898 gene might induce apoptosis and suppress the growth of tumor cells. The colony formation and tumorigenicity experiments are effective for primary function analysis and can provide clues for further researches.