慢病毒介导的靶向干扰PARP-1基因乳腺癌稳定细胞株的构建

Construction of a stable breast cancer cell line for lentivirus-mediated RNA interference to silence PARP-1

Objective To construct a PARP-1 lentivirus interference vector and establish a stably infected human breast cancer MDA-MB-231 cell line. Methods According to genetic information of PARP-1 cDNA sequence provided by GenBank，a targeted PARP-1 lentiviral vector was constructed. Both the lentiviral vector and virus packaging plasmids were transfected into 293T cells to produce recombinant lentivirus，which was infected into the human breast cancer MDA-MB-231 cell line（PARP-1 group）. Negative control and blank control groups were also prepared. The mRNA and protein expression of PARP-1 was measured using real-time quantitative PCR and Western blot. Results PARP-1-RNAi lentiviral vector was constructed and infected into MDA-MB-231 cells. The mRNA expression levels of PARP-1 in blank control group，negative control group and PARP-1 group were，respectively，1.001±0.056，1.022±0.021 and 0.166±0.041； the relative expression levels of PARP-1 protein were 0.963±0.006，0.943±0.008 and 0.573±0.011. Compared with the blank control group and negative control group，the expression of PARP-1 mRNA and protein was significantly lower in the PARP-1 group（P<0.05）. Conclusion A breast cancer cell line stably expressing low levels of PARP-1 has been established.