{"title":"姜黄素诱导人肾细胞癌细胞株ACHN中NF-κB和Bcl-2/Bax的表达","authors":"Gang Li, Tie Chong, Ziming Wang","doi":"10.1016/S1007-4376(09)60087-3","DOIUrl":null,"url":null,"abstract":"<div><h3>Objective</h3><p>To explore the <em>in vitro</em> effects of curcumin on the proliferation and apoptosis of the human renal cell carcinoma cell line ACHN, and to investigate its mechanisms of action.</p></div><div><h3>Methods</h3><p>The human renal cell carcinoma cell line ACHN was treated with different concentrations of curcumin for 24 h. The MTT assay was used to evaluate the cytotoxic effects of curcumin and flow cytometry was utilized to observe and detect the apoptosis of ACHN cells induced by curcumin. The expression levels of Bcl-2, Bax and NF-κBP65 mRNA were evaluated by Reverse Transcription-Polymerase Chain Reaction (RT-PCR), while the expression of Bcl-2, Bax, NF-κBP65 and IκB proteins was evaluated by Western blot.</p></div><div><h3>Results</h3><p>The concentrations of curcumin used significantly inhibited the proliferation of ACHN human renal cell carcinoma cells <em>in vitro</em> in a dose and time-dependent manner (<em>F</em><sub>time</sub> = 5.55, <em>P</em> < 0.05; <em>F</em><sub>dose</sub>=110.05, <em>P</em> < 0.05). Obvious apoptosis of cells treated with different concentrations of curcumin could be observed by FCM. Compared with the control group, the apoptosis rates of curcumin-treated cells were markedly increased (<em>F</em>=96.35, <em>P</em> < 0.05). Lower dose of curcumin significantly induced the apoptosis of ACHN cells. With intervention of different concentrations of curcumin (0, 10, 20 and 40 μmol/L) for 24 h, the expression levels of Bcl-2 and NF-κBP65 mRNA in ACHN cells were decreased while the expression level of Bax mRNA was increased (<em>P</em> < 0.05), and Bcl-2, and NF-κBP65 protein decreased, while Bax and IκB protein increased compared with those in the untreated group.</p></div><div><h3>Conclusion</h3><p>Curcumin inhibited proliferation and increased apoptosis of the human renal cell carcinoma cell line ACHN. These curcumin effects appear to involve up-regulating IκB, down-regulating NF-κB, and regulating the expression of the apoptosis genes Bcl-2/Bax.</p></div>","PeriodicalId":100807,"journal":{"name":"Journal of Nanjing Medical University","volume":"23 6","pages":"Pages 386-391"},"PeriodicalIF":0.0000,"publicationDate":"2009-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/S1007-4376(09)60087-3","citationCount":"3","resultStr":"{\"title\":\"Curcumin induces the expression of NF-κB and Bcl-2/Bax in human renal cell carcinoma cell line ACHN\",\"authors\":\"Gang Li, Tie Chong, Ziming Wang\",\"doi\":\"10.1016/S1007-4376(09)60087-3\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div><h3>Objective</h3><p>To explore the <em>in vitro</em> effects of curcumin on the proliferation and apoptosis of the human renal cell carcinoma cell line ACHN, and to investigate its mechanisms of action.</p></div><div><h3>Methods</h3><p>The human renal cell carcinoma cell line ACHN was treated with different concentrations of curcumin for 24 h. The MTT assay was used to evaluate the cytotoxic effects of curcumin and flow cytometry was utilized to observe and detect the apoptosis of ACHN cells induced by curcumin. The expression levels of Bcl-2, Bax and NF-κBP65 mRNA were evaluated by Reverse Transcription-Polymerase Chain Reaction (RT-PCR), while the expression of Bcl-2, Bax, NF-κBP65 and IκB proteins was evaluated by Western blot.</p></div><div><h3>Results</h3><p>The concentrations of curcumin used significantly inhibited the proliferation of ACHN human renal cell carcinoma cells <em>in vitro</em> in a dose and time-dependent manner (<em>F</em><sub>time</sub> = 5.55, <em>P</em> < 0.05; <em>F</em><sub>dose</sub>=110.05, <em>P</em> < 0.05). Obvious apoptosis of cells treated with different concentrations of curcumin could be observed by FCM. Compared with the control group, the apoptosis rates of curcumin-treated cells were markedly increased (<em>F</em>=96.35, <em>P</em> < 0.05). Lower dose of curcumin significantly induced the apoptosis of ACHN cells. With intervention of different concentrations of curcumin (0, 10, 20 and 40 μmol/L) for 24 h, the expression levels of Bcl-2 and NF-κBP65 mRNA in ACHN cells were decreased while the expression level of Bax mRNA was increased (<em>P</em> < 0.05), and Bcl-2, and NF-κBP65 protein decreased, while Bax and IκB protein increased compared with those in the untreated group.</p></div><div><h3>Conclusion</h3><p>Curcumin inhibited proliferation and increased apoptosis of the human renal cell carcinoma cell line ACHN. These curcumin effects appear to involve up-regulating IκB, down-regulating NF-κB, and regulating the expression of the apoptosis genes Bcl-2/Bax.</p></div>\",\"PeriodicalId\":100807,\"journal\":{\"name\":\"Journal of Nanjing Medical University\",\"volume\":\"23 6\",\"pages\":\"Pages 386-391\"},\"PeriodicalIF\":0.0000,\"publicationDate\":\"2009-11-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://sci-hub-pdf.com/10.1016/S1007-4376(09)60087-3\",\"citationCount\":\"3\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Journal of Nanjing Medical University\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://www.sciencedirect.com/science/article/pii/S1007437609600873\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"\",\"JCRName\":\"\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Journal of Nanjing Medical University","FirstCategoryId":"1085","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S1007437609600873","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 3
摘要
目的探讨姜黄素对人肾细胞癌细胞株ACHN增殖和凋亡的体外影响,并探讨其作用机制。方法不同浓度姜黄素作用于人肾细胞癌细胞株ACHN 24 h,采用MTT法评价姜黄素的细胞毒作用,采用流式细胞术观察并检测姜黄素诱导的ACHN细胞凋亡情况。采用逆转录聚合酶链反应(RT-PCR)检测Bcl-2、Bax和NF-κBP65 mRNA的表达水平,Western blot检测Bcl-2、Bax、NF-κBP65和i -κ b蛋白的表达水平。结果姜黄素对体外培养的人肾细胞癌ACHN细胞增殖有明显的抑制作用,且呈剂量和时间依赖性(Ftime = 5.55, P <0.05;Fdose=110.05, P <0.05)。流式细胞仪观察到不同浓度姜黄素处理的细胞均有明显的凋亡。与对照组相比,姜黄素处理的细胞凋亡率明显升高(F=96.35, P <0.05)。低剂量姜黄素显著诱导ACHN细胞凋亡。不同浓度姜黄素(0、10、20、40 μmol/L)干预24 h后,ACHN细胞中Bcl-2和NF-κBP65 mRNA表达水平降低,Bax mRNA表达水平升高(P <与未治疗组比较,Bcl-2、NF-κBP65蛋白含量降低,Bax、i -κ b蛋白含量升高。结论姜黄素能抑制人肾细胞癌细胞株ACHN的增殖,增加细胞凋亡。姜黄素的这些作用似乎涉及上调i -κB,下调NF-κB,调节凋亡基因Bcl-2/Bax的表达。
Curcumin induces the expression of NF-κB and Bcl-2/Bax in human renal cell carcinoma cell line ACHN
Objective
To explore the in vitro effects of curcumin on the proliferation and apoptosis of the human renal cell carcinoma cell line ACHN, and to investigate its mechanisms of action.
Methods
The human renal cell carcinoma cell line ACHN was treated with different concentrations of curcumin for 24 h. The MTT assay was used to evaluate the cytotoxic effects of curcumin and flow cytometry was utilized to observe and detect the apoptosis of ACHN cells induced by curcumin. The expression levels of Bcl-2, Bax and NF-κBP65 mRNA were evaluated by Reverse Transcription-Polymerase Chain Reaction (RT-PCR), while the expression of Bcl-2, Bax, NF-κBP65 and IκB proteins was evaluated by Western blot.
Results
The concentrations of curcumin used significantly inhibited the proliferation of ACHN human renal cell carcinoma cells in vitro in a dose and time-dependent manner (Ftime = 5.55, P < 0.05; Fdose=110.05, P < 0.05). Obvious apoptosis of cells treated with different concentrations of curcumin could be observed by FCM. Compared with the control group, the apoptosis rates of curcumin-treated cells were markedly increased (F=96.35, P < 0.05). Lower dose of curcumin significantly induced the apoptosis of ACHN cells. With intervention of different concentrations of curcumin (0, 10, 20 and 40 μmol/L) for 24 h, the expression levels of Bcl-2 and NF-κBP65 mRNA in ACHN cells were decreased while the expression level of Bax mRNA was increased (P < 0.05), and Bcl-2, and NF-κBP65 protein decreased, while Bax and IκB protein increased compared with those in the untreated group.
Conclusion
Curcumin inhibited proliferation and increased apoptosis of the human renal cell carcinoma cell line ACHN. These curcumin effects appear to involve up-regulating IκB, down-regulating NF-κB, and regulating the expression of the apoptosis genes Bcl-2/Bax.