|
|
|
| Anticancer effect of SN-38 combined with sorafenib on hepatocellular carcinoma in vitro and its mechanism |
| XU Li, ZHU Yuan-run, CHEN Jian, YANG Xiao-chun, LUO Pei-hua |
| College of Pharmaceutical Sciences, Zhejiang University, Hangzhou 310058, China |
|
|
|
Abstract Objective:To investigate the anticancer effect and its mechanism of SN-38 combined with sorafenib on hepatocellular cancer cell lines HepG-2 and BEL-7402. Methods:SRB colorimetry was employed to measure the viability of HepG-2 and BEL-7402 cells after the treatment of SN-38 with sorafenib. Propidium iodide flow cytometric assay and DAPI staining were used to evaluate the apoptosis of HCC cells. Western blotting was conducted to detect the expression level of apoptosis-related and DNA damage-related proteins. Results:SRB colorimetry showed the synergistic anticancer activities of SN-38 combined with sorafenib, with a combination index of <0.9. The apoptotic rates of HepG-2 cells in control, 60 nmol/L SN-38, 2.5μmol/L sorafenib and combination groups were 4.25%±2.45%, 28.95%±10.75%, 3.49%±2.49% and 53.19%±11.21%, respectively(P<0.05). Western blotting showed that the combination of these two drugs increased the enzymolysis of PARP, Caspase-8 and Caspase-3, and promoted the expression levels of p53, p21 and γ-H2AX significantly. Conclusion:SN-38 and sorafenib have synergistic anticancer activity on hepatocellular carcinoma cells in vitro with the augmentation of apoptosis.
|
|
Received: 15 May 2015
|
|
|
SN-38与索拉非尼联合应用体外抗肝癌效果及其机制
目的:研究伊立替康的活性代谢物SN-38与索拉非尼联合作用于HepG-2和BEL-7402细胞株的抗肝癌效果及其相关机制。方法:利用磺酰罗丹明B显色法测定SN-38与索拉非尼单用或合用后HepG-2和BEL-7402细胞的存活率。利用PI染色结合流式细胞术及DAPI染色法检测细胞凋亡,同时利用蛋白质印迹法检测凋亡相关蛋白、DNA损伤标志蛋白的表达情况。结果:与单药比较,SN-38与索拉非尼合用对细胞的抑制作用增强,合用指数小于0.9。对照组、SN-38组、索拉非尼组、合用组HepG-2细胞凋亡率分别为4.25%±2.45%、28.95%±10.75%、3.49%±2.49%、53.19%±11.21%,合用组细胞凋亡率增加(与其他组比较均P<0.05)。同时合用组凋亡相关蛋白多聚二磷酸腺苷核糖聚合酶(PARP)、半胱氨酸天冬氨酸蛋白酶(Caspase)8、Caspase-3的蛋白酶切量以及p53蛋白、p21蛋白、DNA损伤标志蛋白磷酸化的组蛋白H2AX的表达量均增加。结论:在细胞水平上,SN-38与索拉非尼联合应用能够通过p53表达增加促进肝癌细胞凋亡,因此具有抗肝癌效果。
关键词：
肝肿瘤/药物疗法,
癌,肝细胞/病理学,
苯磺酸盐类/治疗应用,
喜树碱/类似物和衍生物,
喜树碱/治疗应用,
药物疗法,联合,
细胞凋亡,
基因, p53
|
|
| [[1]] |
TORRE L A, BRAY F, SIEGEL R L, et al. Global cancer statistics, 2012[J]. CA Cancer J Clin, 2015,65(2):87-108.
|
|
|
| [[2]] |
XIE B, WANG D H, SPECHLER S J. Sorafenib for treatment of hepatocellular carcinoma:a systematic review[J]. Dig Dis Sci, 2012,57(5):1122-1129.
|
|
|
| [[3]] |
BRANDI G, BIASCO G, MIRARCHI M G, et al. A phase I study of continuous hepatic arterial infusion of irinotecan in patients with locally advanced hepatocellular carcinoma[J]. Dig Liver Dis, 2011,43(12):1015-1021.
|
|
|
| [[4]] |
FONT A, SANCHEZ J M, ROSELL R, et al. Phase I study of weekly CPT-11(irinotecan)/docetaxel in patients with advanced solid tumors[J]. Lung Cancer, 2002,37(2):213-218.
|
|
|
| [[5]] |
HOLLEBECQUE A, MALKA D, FERTÉ C, et al. Systemic treatment of advanced hepatocellular carcinoma:from disillusions to new horizons[J]. Eur J Cancer, 2015,51(3):327-339.
|
|
|
| [[6]] |
BOIGE V, TAÏEB J, HEBBAR M, et al. Irinotecan as first-line chemotherapy in patients with advanced hepatocellular carcinoma:a multicenter phase II study with dose adjustment according to baseline serum bilirubin level[J]. Eur J Cancer, 2006,42(4):456-459.
|
|
|
| [[7]] |
WYLLIE A H. Apoptosis:an overview[J]. Br Med Bull, 1997,53(3):451-465.
|
|
|
| [[8]] |
HENGARTNER M O. The biochemistry of apoptosis[J]. Nature, 2000,407(6805):770-776.
|
|
|
| [[9]] |
RICCARDI C, NICOLETTI I. Analysis of apoptosis by propidium iodide staining and flow cytometry[J]. Nat Protoc, 2006,1(3):1458-1461.
|
|
|
| [[10]] |
NUNEZ R. DNA measurement and cell cycle analysis by flow cytometry[J]. Curr Issues Mol Biol, 2001,3(3):67-70.
|
|
|
| [[11]] |
VOGELSTEIN B, LANE D, LEVINE A J. Surfing the p53 network[J]. Nature, 2000,408(6810):307-310.
|
|
|
| [[12]] |
EVAN G I, VOUSDEN K H. Proliferation, cell cycle and apoptosis in cancer[J]. Nature, 2001,411(6835):342-348.
|
|
|
| [[13]] |
BUNZ F, DUTRIAUX A, LENGAUER C, et al. Requirement for p53 and p21 to sustain G2 arrest after DNA damage[J]. Science, 1998,282(5393):1497-1501.
|
|
|
|
Viewed |
|
|
|
Full text
|
|
|
|
|
Abstract
|
|
|
|
|
Cited |
|
|
|
|
| |
Shared |
|
|
|
|
| |
Discussed |
|
|
|
|
