Cheng Yan, Hasiqi Mei-Ge, Qin Xiao-Zhen, Tang Xiang-You, Chen Jian-Nan, Wang Hui-Yin, Gao Ao
Experimental Center of Northwest University for Nationalities, Lanzhou 730124, China.
Life Science and Engineering College of Northwest University for Nationalities, Lanzhou 730124, China.
Zhongguo Zhong Yao Za Zhi. 2016 Feb;41(4):603-608. doi: 10.4268/cjcmm20160409.
To investigate the effects of Liu Tea extracts(LTE) on proliferation, apoptosis and drug sensitivity of drug-resistant gastric cancer cell BGC823/5-FU. MTT assay was used to analyze effect of LTE on cell growth and sensitivity chemotherapeutic drugs, and synergistic effect of the combination of LTE with 5-FU on BGC823/5-FU cells. Combination index (CI) was calculated by CompuSyn. Cell apoptosis was measured by flow cytometry (FCM). Protein expressions of P-gp, Bcl-2, Bax and Caspase-3 (17KD) were detected by Western blot at different concentrations of LTE in BGC823/5-FU cells (100, 200, 400 mg•L⁻¹). The results showed that LTE had an inhibitory effect on growth of BGC823/5-FU cell in a dose-time-dependent manner and significantly reduced IC₅₀ of 5-FU, CDDP, PTX and ADM to BGC823/5-FU cells(P<0.05), indicating it could reverse tolerance of drug resistant cells to chemotherapeutic drugs, with reversion multiples of 2.35, 1.68, 1.96, 0.52. The combination of LTE with 5-FU had positive synergistic effect on the BGC-823 cell line. FCM assay suggested that LTE could induce BGC823/5-FU apoptosis. The apoptosis rate was up to 46.2% when the cells were treated with 800 mg•L⁻¹ LTE after 24 h(P<0.01). According to the protein detection results, with the increase in concentration of LTE, the protein expression of Bcl-2 was gradually decreased(P<0.01), the expression of Bax and Caspase-3 were extremely increased(P<0.01), with statistical significance in difference(P<0.01) but no difference in the expression of P-gp between experiment group and control group. LTE can inhibit the growth of drug-resistant human gastric cancer cell BGC823/5-FU and reverse its chemotherapeutic tolerance to some extent. Inhibition of antiapoptotic proteins, activation of proapoptotic proteins and induction of apoptosis of resistant cells may be its main mechanisms.
探讨六茶提取物(LTE)对耐药胃癌细胞BGC823/5-FU增殖、凋亡及药敏性的影响。采用MTT法分析LTE对细胞生长及化疗药物敏感性的影响,以及LTE与5-FU联合应用对BGC823/5-FU细胞的协同作用。用CompuSyn计算联合指数(CI)。采用流式细胞术(FCM)检测细胞凋亡情况。在不同浓度(100、200、400 mg•L⁻¹)的LTE作用下,用蛋白质免疫印迹法检测BGC823/5-FU细胞中P-糖蛋白(P-gp)、Bcl-2、Bax和半胱天冬酶-3(17KD)的蛋白表达。结果显示,LTE对BGC823/5-FU细胞生长具有剂量-时间依赖性抑制作用,并显著降低5-FU、顺铂(CDDP)、紫杉醇(PTX)和阿霉素(ADM)对BGC823/5-FU细胞的半数抑制浓度(IC₅₀)(P<0.05),表明其可逆转耐药细胞对化疗药物的耐受性,逆转倍数分别为2.35、1.68、1.96、0.52。LTE与5-FU联合应用对BGC-823细胞系具有正协同作用。FCM检测表明,LTE可诱导BGC823/5-FU细胞凋亡。24 h时,800 mg•L⁻¹ LTE作用后细胞凋亡率高达46.2%(P<0.01)。根据蛋白质检测结果,随着LTE浓度升高,Bcl-2蛋白表达逐渐降低(P<0.01),Bax和Caspase-3表达显著升高(P<0.01),差异具有统计学意义(P<0.01),但实验组与对照组P-gp表达无差异。LTE能够抑制耐药人胃癌细胞BGC823/5-FU生长,并在一定程度上逆转其化疗耐受性。抑制抗凋亡蛋白、激活促凋亡蛋白以及诱导耐药细胞凋亡可能是其主要作用机制。