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Malignant phenotype of PC3 cell line was inhibited by siRNA targeting PAR gene
Xiaofeng Xu1, Siwei Zhou, Zhengyu Zhang
1Department of Urology Surgery, Nanjing General Hospital of Nanjing Military Command, Nanjing 210002, China. shxxf@tom.com
Abstract:
To investigate the effects of down-regulation of prostate androgen regulated (PAR) expression on proliferation of PC3 cells by using RNA interference (RNAi), suppression of PAR expression was achieved by transfection of PC3 cells with short hairpin RNA (shRNA) expression vectors against PAR, designated as psiRNA-PAR1, psiRNA-PAR2 and psiRNA-PAR3. The inhibitory effects were confirmed by RT-PCR. The growth features of PC3 transfectants were analyzed by cell counts, colon formation in soft agar and flow cytometry. The expression of PAR was suppressed by the three shRNA expression vectors. psiRNA-PAR1 was shown to inhibit the PAR expression most efficiently, with the inhibitory rate reaching a peak at (81.18+/-1.68)% 48 h after the transfection. PC3 transfectants exhibited a decreased proliferation in cell culture and a low efficiency of colon formation in soft agar. Flow cytometry revealed a G(2)/M arrest and induced apoptosis. Down-regulated PAR expression inhibited the growth of PC3 cells by inducing G(2)/M arrest and activating apoptotic pathway. As a potential proto-oncogene that triggers and/or has persistent malignant proliferation, PAR may serves as a very target for the gene therapy.
Insights
Down-regulating prostate androgen regulated (PAR) expression using RNA interference inhibits PC3 cell proliferation. This targeted gene therapy approach induces cell cycle arrest and apoptosis, offering a potential strategy for prostate cancer treatment.
Area of Science:
- Oncology
- Molecular Biology
- Gene Therapy
Background:
- Prostate androgen regulated (PAR) protein is implicated in cell proliferation.
- Understanding PAR's role is crucial for developing targeted prostate cancer therapies.
Purpose of the Study:
- To investigate the effects of down-regulating prostate androgen regulated (PAR) expression on PC3 prostate cancer cell proliferation using RNA interference (RNAi).
Main Methods:
- PC3 cells were transfected with short hairpin RNA (shRNA) expression vectors targeting PAR (psiRNA-PAR1, psiRNA-PAR2, psiRNA-PAR3).
- PAR expression levels were confirmed via RT-PCR.
- Cell proliferation was assessed through cell counts, soft agar colony formation assays, and flow cytometry.
Main Results:
- All three shRNA vectors successfully suppressed PAR expression, with psiRNA-PAR1 showing the highest efficiency (81.18% inhibition at 48h).
- PC3 transfectants displayed reduced proliferation and significantly lower colony formation in soft agar.
- Flow cytometry analysis indicated a G2/M phase cell cycle arrest and induced apoptosis in treated cells.
Conclusions:
- Down-regulation of PAR expression effectively inhibits PC3 cell growth by inducing G2/M arrest and activating apoptotic pathways.
- PAR may function as a proto-oncogene promoting malignant proliferation, making it a promising target for gene therapy in prostate cancer.
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