姜黄素抑制肾癌ACHN细胞增殖及促凋亡的实验研究

Effects of curcumin on the proliferation and apoptosis of human renal cell carcinoma cell line ACHN

目的观察姜黄素对人肾癌ACHN细胞株增殖及细胞凋亡的影响,探讨姜黄素诱导ACHN细胞株凋亡的作用机制。方法不同浓度姜黄素作用人肾癌ACHN细胞24 h后,应用MTT比色法检测姜黄素对人肾癌ACHN细胞的增殖抑制率;流式细胞术检测姜黄素诱导细胞的凋亡率;RT-PCR检测姜黄素对ACHN细胞Bcl-2、Bax、NF-κBP65 mRNA表达的影响;Western blot方法检测其对细胞Bcl-2、Bax、NF-κBP65I、κB蛋白表达的影响。结果姜黄素对人肾癌ACHN细胞有明显的抑制作用,可引起细胞凋亡,并且存在剂量和时间依赖;不同浓度姜黄素作用细胞24 h后,Bcl-2、NF-κBP65 mRNA水平下降,Bax mRNA水平升高(P<0.05),Bcl-2、NF-κBP65蛋白表达量下降,BaxI、κB蛋白表达量升高(P<0.05)。结论姜黄素通过上调IκB,下调NF-κB活性,调控凋亡基因Bcl-2/Bax的表达,抑制人肾癌ACHN细胞的增殖,诱导人肾癌ACHN细胞的凋亡。

Objective To explore the effects of curcumin on the proliferation and apoptosis of human renal cell carcinoma cell line ACHN in vitro, and to investigate its mechanisms. Methods The human renal cell carcinoma cell line ACHN was treated with curcumin of different concentrations for 24 h respectively. The MTT assay was used to evaluate the cytotoxic effects of curcumin on ACHN cells. The flow cytometry were utilized to observe and detect the apoptosis of ACHN ceils induced by curcumin. The expression of Bcl-2, Bax and NF-κP65 mRNA were evaluated by Reverse Transcription-Polymerase Chain Reaction （RT-PCR） method. The expression of Bcl-2, Bax, NF-κBP65 and κB were evaluated by Western blot method. Results The curcumin of different concentrations could significantly inhibit the proliferation and induce apoptosis of ACHN cells, with obvious dose- and time-dependent effects. With intervention of different concentrations of curcumin for 24 h, the expression of Bcl-2 and NF-κBP65 mRNA in ACHN cells was decreased while the expression of Bax mRNA was increased （P〈0.05）. Bcl-2 and NF-κBP65 protein decreased, while Bax and κB protein increased, as compared with those in the untreated group （〈~ 0.05）. Conclusion The proliferation-inhibiting effects of curcumin on human renal cell carcinoma cell line ACHN may be related to up-regulating IκB and down-regulating NF-κB, which regulate the expression of the apoptosis genes Bcl-2/Bax and promote the apoptosis.