[Effect of neferine on adriamycin-resistance of thermotolerant hepatocarcinoma cell line HepG2/thermotolerance].
[Effect of neferine on adriamycin-resistance of thermotolerant hepatocarcinoma cell line HepG2/thermotolerance].
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发表时间：
2007-04
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通讯作者：
Xiaohong Ai;Xiao-Qing Tang;Yanping Liu;Hua-qing Liu;Lin Dong
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作者：
Xiaohong Ai;Xiao-Qing Tang;Yanping Liu;Hua-qing Liu;Lin Dong
背景与目的逆转耐热癌细胞的多药耐药（MDR）是目前肿瘤热疗学研究的热点。本研究旨在探讨肝癌耐热细胞系HepG 2/thermotolerance对阿霉素（ADR）的耐药性及甲基莲心碱（Nef）对HepG 2/thermotolerance耐药性的影响。方法MTT法检测细胞增殖; PI染色流式细胞仪检测细胞凋亡。用异硫氰酸荧光素（FITC）标记的抗bcl-2抗体通过流式细胞术测量Bcl-2的表达。结果43 ℃培养24 h，HepG 2/耐热细胞的增殖率和凋亡率分别为（89.6 ± 5.4）%和（13.6 ± 5.4）%，而HepG 2细胞的增殖率和凋亡率分别为（23.9 ± 3.6）%和（68.9 ± 7.3）%。ADR对HepG 2/耐热细胞的50%抑制浓度（IC 50）是HepG 2细胞的10.8倍[（113.7+/-12.7）micromol/L vs.（10.5+/-2.3）micromol/L]。1、10、100 μ mol/L ADR在37 ℃作用24 h后，HepG 2/耐热细胞的凋亡率分别为（9.3 ± 2.6）%、（17.8 ± 7.3）%和（32.9 ± 8.6）%，而HepG 2细胞分别为（14.3 ± 3.9）%、（38.9 ± 6.8）%和（62.7 ± 5.9）%。在10和40 μ mol/L Nef的存在下，ADR对HepG 2/耐热细胞的IC 50显著降低，（113.7+/-12.7）μ mol/L至（63.7+/-5.6）微摩尔/升，（16.8+/-2.8）微摩尔/升，10 μ mol/L ADR诱导的细胞凋亡率分别由（17.8 ± 4.3）%增加到（26.8 ± 5.9）%和（34.9 ± 8.7）%。Bcl-2在HepG 2/热耐受细胞中过表达，而40 μ mol/L Nef处理24 h后Bcl-2表达下调。结论HepG 2/耐热细胞对ADR具有耐药性。Nef可能通过下调Bcl-2的表达而逆转HepG 2/耐热细胞的ADR抗性。
BACKGROUND & OBJECTIVE Nowadays, reversing the multidrug resistance (MDR) of thermotolerant carcinoma cells is a hot topic in tumor thermatology. This study was to investigate the adriamycin (ADR)-resistance of thermotolerant hepatocarcinoma cell line HepG2/thermotolerance and the effect of neferine (Nef) on the ADR-resistance of HepG2/thermotolerance cells. METHODS Cell proliferation was measured by MTT assay. Cell apoptosis was detected by flow cytometry (FCM) with PI staining. The expression of Bcl-2 was measured by FCM using fluorescein isothiocyanate (FITC)-conjugated anti-bcl-2 antibodies. RESULTS The proliferation rate and apoptosis rate of HepG2/thermotolerance cells cultured in 43 degrees C for 24 h were (89.6+/-5.4)% and (13.6+/-5.4)%, respectively; however, those of HepG2 cells were (23.9+/-3.6)% and (68.9+/-7.3)%, respectively. The 50% inhibition concentration (IC50) of ADR was 10.8 times higher for HepG2/thermotolerance cells than for HepG2 cells [(113.7+/-12.7) micromol/L vs. (10.5+/-2.3) micromol/L]. When treated with 1, 10, 100 micromol/L ADR at 37 degrees C for 24 h, the apoptosis rates of HepG2/thermotolerance cells were (9.3+/-2.6)%, (17.8+/-7.3)%, and (32.9+/-8.6)%, respectively, but those of HepG2 cells were (14.3+/-3.9)%, (38.9+/-6.8)%, and (62.7+/-5.9)%, respectively. In the presence of 10 and 40 micromol/L Nef, the IC50 of ADR for HepG2/thermotolerance cells was significantly decreased from (113.7+/-12.7) micromol/L to (63.7+/-5.6) micromol/L and (16.8+/-2.8) micromol/L, and the cell apoptosis induced by 10 micromol/L ADR was significantly increased from (17.8+/-4.3)% to (26.8+/-5.9)% and (34.9+/-8.7)%, respectively. Bcl-2 was overexpressed in HepG2/thermotolerance cells, whereas it was down-regulated when the cells were treated with 40 micromol/L Nef for 24 h. CONCLUSIONS HepG2/thermotolerance cells are ADR-resistant. Nef may reverse the ADR-resistance of HepG2/thermotolerance cells by down-regulating Bcl-2 expression.