姜黄素和顺铂联用对雄激素非依赖性前列腺癌细胞株PC-3的作用
摘要
目的
研究姜黄素和顺铂联用抑制雄激素非依赖性前列腺癌细胞株PC-3的增殖及诱导凋亡的作用。
方法
不同浓度的姜黄素(10μmol/L、25μmol/L、50μmol/L、75μmol/L、100μmol/L)和顺铂(3 mg/L)分别和联合作用于雄激素非依赖性前列腺癌细胞株PC-3,MTT比色法检测细胞生长活性,透射电镜观察细胞超微结构变化。流式细胞仪测定细胞周期时相变化及检测细胞凋亡。
结果
姜黄素显著抑制PC-3细胞的生长,呈剂量和时间依赖性。不同浓度姜黄素组之间与不同作用时间组之间的差异均有显著性意义(P均<0.01);顺铂联合不同浓度姜黄素可显著抑制PC-3细胞增殖(P均<0.05),差异有显著性意义。50μmol/L姜黄素和3 mg/L顺铂联合作用PC-3细胞24h后,细胞出现凋亡形态学改变。各浓度姜黄素组作用PC-3细胞24h后,凋亡细胞比例均显著高于空白对照组(P均<0.05),差异有显著性意义, PC-3细胞出现剂量依赖性G2/M期阻滞(P<0.01),姜黄素及顺铂均可诱导PC-3细胞凋亡。两种药物联用后,诱导细胞凋亡作用增强。
结论
姜黄素通过诱导前列腺癌细胞株PC-3的凋亡从而抑制肿瘤细胞的生长。姜黄素与顺铂能够协同抑制人前列腺癌细胞株PC-3的增殖,两种药物联用有增效的作用。
Objective: To study the growth inhibition effects of Curcumin and Cisplatin on androgen independent prostate cancer cell line PC-3.
Methods: The growth activities of cancer cells were studied by MTT colorimetry. Cell cycle phases and the apoptosis were inspected by flowcy-tometery (FCM). The morphological changes of cancer cells were observed under electronic microscopy.
Results: Curcumin obviously suppressed the proliferation of PC-3 cell line in does-dependent and time-dependent manners in vitro. There was an obvious difference of inhibition rate in different concentrations and time groups (P<0.05). Curcumin could lead to characteristic morphological changes of apoptosis in PC-3 cell line after 24 hours. The ratios of apoptosis cell were significantly higher than that of controls (P<0.05) after curcumin influence the PC-3 cell line. Curcumin could arrest the cell cycle of PC-3 cell line at G2/M phase in does-dependent manners (P<0.01). Thses effects could enhance significantly by cisplatin. The curcumin and cisplatin can all induce cell line PC-3 apoptosis. When the two drugs combined, the inductive effect for cell apoptosis became obvious.
Conclusion: Curcumine could suppress the growth of androgen-indenpent prostate cancer cell lines and promote their apoptosis by different mechanism. Cisplatin can enhance antiproliferative effect of curcumin to hunman prostate cancer cell line PC-3.
研究姜黄素和顺铂联用抑制雄激素非依赖性前列腺癌细胞株PC-3的增殖及诱导凋亡的作用。
方法
不同浓度的姜黄素(10μmol/L、25μmol/L、50μmol/L、75μmol/L、100μmol/L)和顺铂(3 mg/L)分别和联合作用于雄激素非依赖性前列腺癌细胞株PC-3,MTT比色法检测细胞生长活性,透射电镜观察细胞超微结构变化。流式细胞仪测定细胞周期时相变化及检测细胞凋亡。
结果
姜黄素显著抑制PC-3细胞的生长,呈剂量和时间依赖性。不同浓度姜黄素组之间与不同作用时间组之间的差异均有显著性意义(P均<0.01);顺铂联合不同浓度姜黄素可显著抑制PC-3细胞增殖(P均<0.05),差异有显著性意义。50μmol/L姜黄素和3 mg/L顺铂联合作用PC-3细胞24h后,细胞出现凋亡形态学改变。各浓度姜黄素组作用PC-3细胞24h后,凋亡细胞比例均显著高于空白对照组(P均<0.05),差异有显著性意义, PC-3细胞出现剂量依赖性G2/M期阻滞(P<0.01),姜黄素及顺铂均可诱导PC-3细胞凋亡。两种药物联用后,诱导细胞凋亡作用增强。
结论
姜黄素通过诱导前列腺癌细胞株PC-3的凋亡从而抑制肿瘤细胞的生长。姜黄素与顺铂能够协同抑制人前列腺癌细胞株PC-3的增殖,两种药物联用有增效的作用。
Objective: To study the growth inhibition effects of Curcumin and Cisplatin on androgen independent prostate cancer cell line PC-3.
Methods: The growth activities of cancer cells were studied by MTT colorimetry. Cell cycle phases and the apoptosis were inspected by flowcy-tometery (FCM). The morphological changes of cancer cells were observed under electronic microscopy.
Results: Curcumin obviously suppressed the proliferation of PC-3 cell line in does-dependent and time-dependent manners in vitro. There was an obvious difference of inhibition rate in different concentrations and time groups (P<0.05). Curcumin could lead to characteristic morphological changes of apoptosis in PC-3 cell line after 24 hours. The ratios of apoptosis cell were significantly higher than that of controls (P<0.05) after curcumin influence the PC-3 cell line. Curcumin could arrest the cell cycle of PC-3 cell line at G2/M phase in does-dependent manners (P<0.01). Thses effects could enhance significantly by cisplatin. The curcumin and cisplatin can all induce cell line PC-3 apoptosis. When the two drugs combined, the inductive effect for cell apoptosis became obvious.
Conclusion: Curcumine could suppress the growth of androgen-indenpent prostate cancer cell lines and promote their apoptosis by different mechanism. Cisplatin can enhance antiproliferative effect of curcumin to hunman prostate cancer cell line PC-3.
引文
[1] 顾方六主编.现代泌尿肿瘤外科学[M].北京:科学出版社,2003.5.
[2] Kuttan R. Potential anticancer activity of turmeric (curcum a longa) [J]. Cancer Lett, 1985; 29:197.
[3] Lin JK, Jin-shiau SY. Mechanisms of cancer chemoprevention bycurcumi[J].Proc Natl Sci Counc ROC(B),2001,25(2):59-66
[4] Jiang MC, Yang-Yen HF, Yen JJ, et al. Curcumin induces apoptosis in immortalized NIH 3T3 and malignant cancer cell lines. Nutr Cancer. 1996; 26(1):111-20.
[5] Koo JY, Kim HJ, Jung KO, et al. Curcumin inhibits the growth of AGS human gastric carcinoma cells in vitro and shows synergism with 5-fluorouracil. J Med Food, 2004, 7(2): 117-121.
[6] Aggarwal S, Takada Y, Singh S, et al. Inhibition of growth and survival of human head and neck squamous cell carcinoma cells by curcumin via modulation of nuclear factor-kappaB signaling. Int J Cancer, 2004, 111(5): 679-692.
[7] Radhakrishna Pillai G, Srivastava AS, Hassanein TI, et al. Induction of apoptosis in human lung cancer cells by curcumin. Cancer Lett, 2004, 208(2): 163-170.
[8] Han SS, Keum YS, Seo HJ, et al. Curcumin suppresses activation of NF-kappaB and AP-1 induced by phorbol ester in cultured human promyelocytic leukemia cells. J Biochem Mol Biol, 2002, 35(3): 337-342.
[9] 韩锐,主编。肿瘤化学预防及药物治疗[M]。北京:北京医科大学中国协和医科大学联合出版社,1995:418-419
[10] ShaoZhi-Ming, ShenZhen-Zhou, LiuCan-Hui. Curcumin exerts multiple suppressive effects on human breast carcinoma cell[J]. Int J cancer, 2002, 98(2):234.
[11] 吴裕丹,陈燕,陈文娟,等。INF-γ对姜黄素HL-60细胞增殖的影响,[J]。癌症,1999,18:635-638.
[12] Huggins C, Hodges CV. Studies on prostatic cancer. I. The effect of castration, of estrogen and of androgen injection on serum phosphatases in metastatic carcinoma of the prostate.1941[.J]. J urol. 2002 Feb; 167(2 Pt 2):948-51; discussion 952.
[13] Crawford ED,Eisenberger MA, McLeod DG, et al. A controlled trial of leuprolide with and without flutamide in prostatic carcinoma[J].New Engl J Med, 1989 321:419.
[14] Mcleod D.Hormonal therapy in the treatment of carcinoma of the prostate [J].Cancer, 1995, 75:1914-1919.
[15] Plasencia C, Dayam R, Wang Q, et al. Discovery and preclinical evaluation of a novel class of small-molecule compounds in hormone-dependent and -independent cancer cell lines[J].Mol Cancer Ther. 2005 Jul; 4(7):1105-13.
[16] Poindessous V, Koeppel F, et al. Marked activity of irofulven toward human carcinoma cells: comparison with cisplatin and ecteinascidin[J].Clin Cancer Res. 2003 Jul; 9(7):2817-25.
[17] Shenouda NS, Zhou C, Browning JD, et al. Phytoestrogens in common herbs regulates prostate cancer cell growth in vitro[J]. Nutr Cancer. 2004; 49(2):200-8.
[18] Choudhuri T, Pal S, Agwarwal ML, et al. Curcumin induces apoptosis in human breast cancer cells through p53-dependent Bax induction[J]. FEBS Lett. 2002 Feb 13; 512(1-3):334-40.
[19] Han SS, Chung ST, Robertson DA, et al. Curcumin causes the growth arrest and apoptosis of B cell lymphoma by downregulation of egr-1, c-myc, bcl-XL, NF-kappa B, and p53[J]. Clin Immunol. 1999 Nov; 93(2):152-61.
[20] Anto RJ, Mukhopadhyay A, Denning K, et al. Curcumin (diferuloylmethane) induces apoptosis through activation of caspase-8, BID cleavage and cytochrome c release: its suppression by ectopic expression of Bcl-2 and Bcl-xl. Carcinogenesis. 2002 Jan; 23(1):143-50.
[21] Chen H, Zhang ZS, Zhang YL, et al. Curcumin inhibits cell proliferation by interfering with the cell cycle and inducing apoptosis in colon carcinoma cells[J]. Anticaner Res, 1999, 19(5A):3675-3680.
[22] Mizutani Y, Wu XX, et al. Chemoimmunosen-sitizition of the T24 human bladder cancer line to Fas-mediated cytotoxicity and apoptosis by cisplatin and 5-flurouracial[J]. Oncol Rep, 1999, 6:979-982. .
[1] Small EJ, Reese DM, Vogelzang NJ. Hormone-refractory prostate cancer: an evolving standard of care. Semin Oncol 1999, 26 (5 suppl 17): 6767
[2] Kantoff PW, Halabi S, Conaway M, et al. Hydrocortisone with or without mitoxantrone in men with hormone-refractory prostate cancer: results of the cancer and leukemia group B 9182 study. J Clin Oncol. 1999 Aug;17(8):2506-13.
[3] Ernst DS, Tannock IF, Winquist EW, et al. Randomized, double-blind, controlled trial of mitoxantrone/prednisone and clodronate versus mitoxantrone/prednisone and placebo in patients with hormone-refractory prostate cancer and pain. J Clin Oncol. 2003 Sep 1; 21(17):3335-42.
[4] Hudes G, Einhorn L, Ross E, et al. Vinblastine versus vinblastine plus oral estramustine phosphate for patients with hormone-refractory prostate cancer: A Hoosier Oncology Group and Fox Chase Network phase III trial. J Clin Oncol. 1999 Oct; 17(10):3160-6.
[5] Khan MA, Carducci MA, Partin AW, et al. The evolving role of docetaxel in the management of androgen independent prostate cancer. J Urol, 2003 Nov; 170(5):1709-16.
[6] Tannock IF, de Wit R, Berry WR, et al. Docetaxel plus prednisone or mitoxantrone plus prednisone for advanced prostate cancer. N Engl J Med, 2004 Oct 7; 351(15):1502-12.
[7] Petrylak DP, Tangen CM, Hussain MH, et al. Docetaxel and estramustine compared with mitoxantrone and prednisone for advanced refractory prostate cancer. N Engl J Med. 2004 Oct 7; 351(15):1513-20.
[8] Walczak JR, Carducci MA; Eastern Cooperative Oncology Group E1899. Phase 3 randomized trial evaluating second-line hormonal therapy versus docetaxel-estramustine combination chemotherapy on progression-free survival in asymptomatic patients with a rising prostate-specific antigen level after hormonal therapy for prostate cancer: an Eastern Cooperative Oncology Group (E1899), Intergroup/Clinical Trials Support Unit study. Urology, 2003 Dec 29;62 Suppl 1:141-6.
[9] Beer TM, Eilers KM, Garzotto M, et al. Quality of life and pain relief during treatment with calcitriol and docetaxel in symptomatic metastatic androgen-independent prostate carcinoma. Cancer, 2004 Feb 15; 100(4):758-63
[10] Dahut WL, Gulley JL, Arlen PM, et al. Randomized phase II trial of docetaxel plus thalidomide in androgen-independent prostate cancer.J Clin Oncol, 2004 Jul 1;22(13):2532-9.
[11] Vordos D, Paule B, Vacherot F, et al. Docetaxel and zoledronic acid in patients with metastatic hormone-refractory prostate cancer. BJU Int, 2004 Sep; 94(4):524-7.
[12] Vogelzang NJ, Karrison T, Stadler WM, et al. A Phase II trial of suramin monthly x 3 for hormone-refractory prostate carcinoma. Cancer. 2004 Jan 1; 100(1):65-71.
[13] Smith MR, Kaufman D, Oh W, et al. Vinorelbine and estramustine in androgen-independent metastatic prostate cancer: a phase II study. Cancer. 2000 Oct 15; 89(8):1824-8.
[14] Di Lorenzo G, Pizza C, Autorino R, et al. Weekly docetaxel and vinorelbine (VIN-DOX) as first line treatment in patients with hormone refractory prostate cancer. Eur Urol. 2004 Dec; 46(6):712-6
[2] Kuttan R. Potential anticancer activity of turmeric (curcum a longa) [J]. Cancer Lett, 1985; 29:197.
[3] Lin JK, Jin-shiau SY. Mechanisms of cancer chemoprevention bycurcumi[J].Proc Natl Sci Counc ROC(B),2001,25(2):59-66
[4] Jiang MC, Yang-Yen HF, Yen JJ, et al. Curcumin induces apoptosis in immortalized NIH 3T3 and malignant cancer cell lines. Nutr Cancer. 1996; 26(1):111-20.
[5] Koo JY, Kim HJ, Jung KO, et al. Curcumin inhibits the growth of AGS human gastric carcinoma cells in vitro and shows synergism with 5-fluorouracil. J Med Food, 2004, 7(2): 117-121.
[6] Aggarwal S, Takada Y, Singh S, et al. Inhibition of growth and survival of human head and neck squamous cell carcinoma cells by curcumin via modulation of nuclear factor-kappaB signaling. Int J Cancer, 2004, 111(5): 679-692.
[7] Radhakrishna Pillai G, Srivastava AS, Hassanein TI, et al. Induction of apoptosis in human lung cancer cells by curcumin. Cancer Lett, 2004, 208(2): 163-170.
[8] Han SS, Keum YS, Seo HJ, et al. Curcumin suppresses activation of NF-kappaB and AP-1 induced by phorbol ester in cultured human promyelocytic leukemia cells. J Biochem Mol Biol, 2002, 35(3): 337-342.
[9] 韩锐,主编。肿瘤化学预防及药物治疗[M]。北京:北京医科大学中国协和医科大学联合出版社,1995:418-419
[10] ShaoZhi-Ming, ShenZhen-Zhou, LiuCan-Hui. Curcumin exerts multiple suppressive effects on human breast carcinoma cell[J]. Int J cancer, 2002, 98(2):234.
[11] 吴裕丹,陈燕,陈文娟,等。INF-γ对姜黄素HL-60细胞增殖的影响,[J]。癌症,1999,18:635-638.
[12] Huggins C, Hodges CV. Studies on prostatic cancer. I. The effect of castration, of estrogen and of androgen injection on serum phosphatases in metastatic carcinoma of the prostate.1941[.J]. J urol. 2002 Feb; 167(2 Pt 2):948-51; discussion 952.
[13] Crawford ED,Eisenberger MA, McLeod DG, et al. A controlled trial of leuprolide with and without flutamide in prostatic carcinoma[J].New Engl J Med, 1989 321:419.
[14] Mcleod D.Hormonal therapy in the treatment of carcinoma of the prostate [J].Cancer, 1995, 75:1914-1919.
[15] Plasencia C, Dayam R, Wang Q, et al. Discovery and preclinical evaluation of a novel class of small-molecule compounds in hormone-dependent and -independent cancer cell lines[J].Mol Cancer Ther. 2005 Jul; 4(7):1105-13.
[16] Poindessous V, Koeppel F, et al. Marked activity of irofulven toward human carcinoma cells: comparison with cisplatin and ecteinascidin[J].Clin Cancer Res. 2003 Jul; 9(7):2817-25.
[17] Shenouda NS, Zhou C, Browning JD, et al. Phytoestrogens in common herbs regulates prostate cancer cell growth in vitro[J]. Nutr Cancer. 2004; 49(2):200-8.
[18] Choudhuri T, Pal S, Agwarwal ML, et al. Curcumin induces apoptosis in human breast cancer cells through p53-dependent Bax induction[J]. FEBS Lett. 2002 Feb 13; 512(1-3):334-40.
[19] Han SS, Chung ST, Robertson DA, et al. Curcumin causes the growth arrest and apoptosis of B cell lymphoma by downregulation of egr-1, c-myc, bcl-XL, NF-kappa B, and p53[J]. Clin Immunol. 1999 Nov; 93(2):152-61.
[20] Anto RJ, Mukhopadhyay A, Denning K, et al. Curcumin (diferuloylmethane) induces apoptosis through activation of caspase-8, BID cleavage and cytochrome c release: its suppression by ectopic expression of Bcl-2 and Bcl-xl. Carcinogenesis. 2002 Jan; 23(1):143-50.
[21] Chen H, Zhang ZS, Zhang YL, et al. Curcumin inhibits cell proliferation by interfering with the cell cycle and inducing apoptosis in colon carcinoma cells[J]. Anticaner Res, 1999, 19(5A):3675-3680.
[22] Mizutani Y, Wu XX, et al. Chemoimmunosen-sitizition of the T24 human bladder cancer line to Fas-mediated cytotoxicity and apoptosis by cisplatin and 5-flurouracial[J]. Oncol Rep, 1999, 6:979-982. .
[1] Small EJ, Reese DM, Vogelzang NJ. Hormone-refractory prostate cancer: an evolving standard of care. Semin Oncol 1999, 26 (5 suppl 17): 6767
[2] Kantoff PW, Halabi S, Conaway M, et al. Hydrocortisone with or without mitoxantrone in men with hormone-refractory prostate cancer: results of the cancer and leukemia group B 9182 study. J Clin Oncol. 1999 Aug;17(8):2506-13.
[3] Ernst DS, Tannock IF, Winquist EW, et al. Randomized, double-blind, controlled trial of mitoxantrone/prednisone and clodronate versus mitoxantrone/prednisone and placebo in patients with hormone-refractory prostate cancer and pain. J Clin Oncol. 2003 Sep 1; 21(17):3335-42.
[4] Hudes G, Einhorn L, Ross E, et al. Vinblastine versus vinblastine plus oral estramustine phosphate for patients with hormone-refractory prostate cancer: A Hoosier Oncology Group and Fox Chase Network phase III trial. J Clin Oncol. 1999 Oct; 17(10):3160-6.
[5] Khan MA, Carducci MA, Partin AW, et al. The evolving role of docetaxel in the management of androgen independent prostate cancer. J Urol, 2003 Nov; 170(5):1709-16.
[6] Tannock IF, de Wit R, Berry WR, et al. Docetaxel plus prednisone or mitoxantrone plus prednisone for advanced prostate cancer. N Engl J Med, 2004 Oct 7; 351(15):1502-12.
[7] Petrylak DP, Tangen CM, Hussain MH, et al. Docetaxel and estramustine compared with mitoxantrone and prednisone for advanced refractory prostate cancer. N Engl J Med. 2004 Oct 7; 351(15):1513-20.
[8] Walczak JR, Carducci MA; Eastern Cooperative Oncology Group E1899. Phase 3 randomized trial evaluating second-line hormonal therapy versus docetaxel-estramustine combination chemotherapy on progression-free survival in asymptomatic patients with a rising prostate-specific antigen level after hormonal therapy for prostate cancer: an Eastern Cooperative Oncology Group (E1899), Intergroup/Clinical Trials Support Unit study. Urology, 2003 Dec 29;62 Suppl 1:141-6.
[9] Beer TM, Eilers KM, Garzotto M, et al. Quality of life and pain relief during treatment with calcitriol and docetaxel in symptomatic metastatic androgen-independent prostate carcinoma. Cancer, 2004 Feb 15; 100(4):758-63
[10] Dahut WL, Gulley JL, Arlen PM, et al. Randomized phase II trial of docetaxel plus thalidomide in androgen-independent prostate cancer.J Clin Oncol, 2004 Jul 1;22(13):2532-9.
[11] Vordos D, Paule B, Vacherot F, et al. Docetaxel and zoledronic acid in patients with metastatic hormone-refractory prostate cancer. BJU Int, 2004 Sep; 94(4):524-7.
[12] Vogelzang NJ, Karrison T, Stadler WM, et al. A Phase II trial of suramin monthly x 3 for hormone-refractory prostate carcinoma. Cancer. 2004 Jan 1; 100(1):65-71.
[13] Smith MR, Kaufman D, Oh W, et al. Vinorelbine and estramustine in androgen-independent metastatic prostate cancer: a phase II study. Cancer. 2000 Oct 15; 89(8):1824-8.
[14] Di Lorenzo G, Pizza C, Autorino R, et al. Weekly docetaxel and vinorelbine (VIN-DOX) as first line treatment in patients with hormone refractory prostate cancer. Eur Urol. 2004 Dec; 46(6):712-6