野生型p53基因cDNA转染对卵巢癌细胞系SKOV-3生物学行为的影响

Biologic Effects of Introduction of WildType p53 cDNA into a Human Ovarian Cancer Cell Line SKOV3

目的：研究野生型ｐ５３基因ｃＤＮＡ转染对卵巢癌细胞系ＳＫＯＶ－３生物学行为的影响。方法：用脂质体介导的转染技术，将含有全长人野生型ｐ５３ｃＤＮＡ的真核表达重组质粒和空载体质粒，分别导入不表达ｐ５３的ＳＫＯＶ－３细胞，观察细胞生物学行为的改变。结果：（１）共获２个ｐ５３ｃＤＮＡ转染克隆（ｐＣ５３）和２个空载体转染克隆（ｐＮｅｏ）；（２）ｐＣ５３和ｐＮｅｏ细胞形态学与ＳＫＯＶ－３无显著差别；（３）ｐＣ５３细胞生长明显比ＳＫＯＶ－３慢，而ｐＮｅｏ细胞生长与ＳＫＯＶ－３相当；（４）ｐＣ５３在软琼脂中形成集落数显著少于ＳＫＯＶ－３细胞和ｐＮｅｏ细胞；（５）ｐＣ５３Ｇ１／Ｇ０期细胞百分比高于ＳＫＯＶ－３和ｐＮｅｏ。结论：野生型ｐ５３ｃＤＮＡ可作为卵巢癌基因治疗的目的基因之一。

Objective: To study the effects on biologic behavior in cells obtained from human ovarian cancer cell line SKOV3 into which the wildtype p53 cDNA was introduced. Method: Recombinant eukaryotic expression vector pC53SN3 containing fulllength human wildtype p53 cDNA and vector containing neomycin resistance gene only were introduced by lipofectaminemediated gene transfection into SKOV3 cell line which does not express endogenous p53. The clones obtained were observed for their biologic behavior. Results: (1) 2 clones named pC53 and 2 clones named pNeo were obtained after pC53SN3 and vector transfection respectively; (2) The morphology of cells either from pC53 or from pNeo did not change significantly with respect to their parental SKOV3; (3) The growth rate of cells from pC53 was much slower than that from SKOV3, while the cell growth curve of pNeo was similar to that of SKOV3; (4) The number of colones formed in the softagar by pC53 was significantly less than that by SKOV3 or by pNeo; (5) The percentage of phase G1/G0 of pC53 was much higher than that of SKOV3 and pNeo. Conclusion: Wildtype p53 cDNA may be considered as one of the target genes for the gene therapy of ovarian cancer.