前列腺癌特异抗原3对前列腺癌LNCaP细胞增殖的影响

摘要

Background and purpose: The increased and specific expression of prostate cancer antigen 3 (PCA3), as one of long non-coding RNA, has been observed in prostate cancer, suggesting that PCA3 may contribute to the development of prostate cancer. Based on these evidence, we propose a series of experiments to primarily investigate the effect of PCA3 on proliferation of human prostate cancer cell line (LNCaP). Methods: PCA3 siRNA was transfected into human prostate cancer LNCaP cells by LipofectamineTM 2000, the transfection efficiency was obeserved by reverse fluorescence microscope, and inhibition of PCA3 by small interfering RNA (siRNA) was quantifed by real time quantitative PCR. The ability of proliferation of LNCaP cells after transfection was analyzed by CCK-8 assay and colony formation assay. Results: The transfection efficiency was about 50%-70% after 48 h following transfection. Compared with PCA3 siRNA (si731) and PCA3 siRNA (si2060), PCA3 siRNA (si 164) has the largest effection on inhibition of PCA3. The ability of proliferation of LNCaP cells in group treated with PCA3 siRNA (si164) was significantly decreased as compared with negative group (P<0.05). Conclusion: PCA3 siRNA (si164) has been proved not only to be effective for down-regulation of PCA3, but also significantly inhibit the proliferation of LNCaP cells, indicating that PCA3 may contribute to the progression of prostate cancer by regulating proliferation and may be a potential, novel drug target against prostate cancer.%背景与目的:前列腺癌抗原3(prostate cancer antigen 3,PCA3)作为一种长链非编码RNA已被证实在前列腺癌中特异性高表达,预示其可能在前列腺癌的发生、发展中发挥重要作用.本研究拟通过靶向干扰PCA3以初步观察其对前列腺癌细胞株LNCaP细胞增殖的影响.方法:使用脂质体介导小干扰RNA(siRNA)转染前列腺癌LNCaP细胞,荧光倒置显微镜观察转染效率,实时定量PCR检测siRNA对PCA3的抑制程度；CCK-8法和克隆形成实验检测转染前列腺癌LNCaP细胞后细胞的增殖能力.结果:荧光倒置显微镜观察转染后48 h转染效率为50％～70％；实时定量PCR检测结果发现,与PCA3 siRNA(si731)和PCA3 siRNA (si2060)比较,PCA3siRNA (si164)对PCA3的抑制程度最大；细胞增殖实验显示,转染PCA3 siRNA (si164)组中LNCaP细胞增殖能力较阴性对照组明显下降(P＜0.05).结论:PCA3 siRNA (si164)在前列腺癌LNCaP细胞株中能靶向沉默PCA3的表达并抑制前列腺癌细胞增殖,表明PCA3通过调节细胞增殖促进前列腺癌发展,并可成为前列腺癌新的治疗靶点.