Cytotoxic effects of bromelain in human gastrointestinal carcinoma cell lines (MKN45, KATO-III, HT29-5F12, and HT29-5M21).
Cytotoxic effects of bromelain in human gastrointestinal carcinoma cell lines (MKN45, KATO-III, HT29-5F12, and HT29-5M21).
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DOI：
10.2147/ott.s43072
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发表时间：
2013
影响因子：
4
通讯作者：
Morris DL
中科院分区：
文献类型：
--
作者：
Amini A;Ehteda A;Masoumi Moghaddam S;Akhter J;Pillai K;Morris DL
菠萝蛋白酶是一种菠萝茎提取物，由于与许多不同的生物过程相互作用而具有多种治疗益处。几项临床前研究和轶事临床观察报告了菠萝蛋白酶的抗癌特性。在本研究中，我们研究了菠萝蛋白酶在四种人胃肠道癌细胞系中的细胞毒性作用及其机制。胃癌细胞系（KATO-III和MKN 45）和HT 29结肠腺癌细胞系的两个化学抗性亚群（HT 29 - 5 M21和HT 29 - 5 F12）用一系列浓度的菠萝蛋白酶处理，以及用顺铂作为阳性对照。在处理72小时后，使用磺酰罗丹明B测定法测定菠萝蛋白酶对癌细胞生长和增殖的影响。Western blotting分析菠萝蛋白酶处理后MKN 45细胞凋亡相关蛋白的表达。来自我们的磺酰罗丹明B测定的数据显示，菠萝蛋白酶抑制HT 29 - 5 F12、HT 29 - 5 M21、MKN 45和KATO-III细胞的增殖，其半最大抑制浓度值分别为29、34、94和142 μg/mL。分析菠萝蛋白酶处理的MKN 45细胞中促凋亡和抗凋亡蛋白的表达，我们观察到半胱天冬酶系统的激活、PARP和p53的裂解、细胞色素C的过表达、磷酸化Akt和Bcl 2的衰减以及MUC 1的去除。除了观察到的半胱天冬酶依赖性细胞凋亡外，裂解的p53的出现支持p53的直接的核凋亡功能。此外，中断Akt信号传导和Bcl 2和MUC 1癌蛋白的衰减表明癌细胞的存活受损。我们的研究结果共同表明，菠萝蛋白酶发挥细胞毒性作用，在一组人胃癌和结肠癌细胞。我们对MKN 45细胞的研究表明菠萝蛋白酶诱导的细胞死亡有不同的机制。菠萝蛋白酶在参与半胱天冬酶系统和细胞核p53的情况下促进细胞凋亡的同时，似乎还通过阻断Akt途径和减弱Bcl 2和MUC 1癌蛋白来损害癌细胞存活。
Bromelain is a pineapple stem extract with a variety of therapeutic benefits arising from interaction with a number of different biological processes. Several preclinical studies and anecdotal clinical observations have reported the anticancer properties of bromelain. In the present study, we investigated the cytotoxic effects of bromelain in four human cancer cell lines of gastrointestinal origin and the mechanisms involved. The gastric carcinoma cell lines (KATO-III and MKN45) and two chemoresistant subpopulations of the HT29 colon adenocarcinoma cell line (HT29-5M21 and HT29-5F12) were treated with a range of concentrations of bromelain, as well as with cisplatin as a positive control. The effect of bromelain on the growth and proliferation of cancer cells was determined using a sulforhodamine B assay after 72 hours of treatment. Expression of apoptosis-associated proteins in MKN45 cells treated with bromelain was analyzed by Western blotting. Data from our sulforhodamine B assay showed that bromelain inhibited proliferation of HT29-5F12, HT29-5M21, MKN45, and KATO-III cells, with respective half maximal inhibitory concentration values of 29, 34, 94, and 142 μg/mL. Analyzing the expression of proapoptotic and antiapoptotic proteins in bromelain-treated MKN45 cells, we observed activation of the caspase system, cleavage of PARP and p53, overexpression of cytochrome C, attenuation of phospho-Akt and Bcl2, and removal of MUC1. Apart from the caspase-dependent apoptosis observed, emergence of cleaved p53 supports a direct, extranuclear apoptotic function of p53. Moreover, interrupted Akt signaling and attenuation of Bcl2 and MUC1 oncoproteins suggest impaired survival of cancer cells. Our findings collectively indicate that bromelain exerts cytotoxic effects in a panel of human gastric and colon carcinoma cells. Our study of MKN45 cells implicated different mechanisms in bromelain-induced cell death. While promoting apoptosis with involvement of the caspase system and extranuclear p53, bromelain also appears to impair cancer cell survival by blocking the Akt pathway and attenuating Bcl2 and MUC1 oncoproteins.