p53反义RNA对肠癌细胞恶性表型的抑制作用

研究ｐ５３反义ＲＮＡ对大肠癌细胞中突变型ｐ５３致癌性的抑制效应,为肿瘤基因治疗提供新思路。方法２．１ｋｂ的人ｐ５３全长ｃＤＮＡ反向插入哺乳动物表达载体ｐＲＥＰ９,得到ｐ５３反义ＲＮＡ表达载体ｐＲＥＰ９－ｐ５３（ＡＳ）,并将其导入人肠癌细胞株ＳＷ１１１６（内源性突变型ｐ５３）,采用ＭＴＴ和ＦＣＭ方法测定ｐＲＥＰ９－ｐ５３（ＡＳ）表达的ｐ５３反义ＲＮＡ对ＳＷ１１１６细胞生长的影响。结果带有ｐＲＥＰ９－ｐ５３（ＡＳ）的ＳＷ１１１６细胞,与对照组ＳＷ１１１６细胞和带ｐＲＥＰ９空载体的ＳＷ１１１６细胞相比,由于ｐ５３反义ＲＮＡ的表达,其增殖速度显著下降,ＦＣＭ的结果也证明带ｐＲＥＰ９－ｐ５３（ＡＳ）的细胞部分受阻于Ｇ０／１期,而带ｐＲＥＰ９空载体的细胞则无明显变化。结论ｐ５３反义ＲＮＡ可以有效地抑制大肠癌细胞中突变型ｐ５３的致癌性,可用于实验性肿瘤基因治疗研究

Inhibition effect of p53 antisense RNA on malignant phenotype of colorectal cancer cells

Objective Inhibition effect ofp53 antisense RNA on malignant phenotype of colorec-tal cancer cells wals studied.Methods For the purpose of inhibiting tumorigenecity of mutant p53 gene in colorectal cancer cells,a 2.1kb human p53 full length cDNA was inserted into a mammalian expression vector pREP9 to make a p53 antisense RNA expression vector pREP9-53(AS).pREP9-p53(AS) was then introduced into human colorectal cancer cell line SW1116(with mutated endoge-nous p53). MTT method and FCM analysis were performed to measure the effect of p53 antisense RNA expressed by pREP9-p53(AS)on SW1116 cell cycle progression.Results and conclusion It was shown that the growth rate of SW1116 cells with pREP-p53(AS) was significantly suppressed by the cxpressionof p53 antisense RNA as compared to the control SW1116 cells andSW1116 cells with pREP9 vector alone.FCM analysis showed that SW1116 cells with pREP-p53(AS) were ar-rested at C o/1 phase,whereas no significant influence can be observed on control SW1116 cells with pREP9.