Madempudi Ratna Sudha, Kalle Arunasree M
a Centre for Research and Development , Unique Biotech Ltd , Hyderabad , India.
b Department of Animal Biology, School of Life Sciences , University of Hyderabad , Hyderabad , India.
Nutr Cancer. 2017 Oct;69(7):1062-1068. doi: 10.1080/01635581.2017.1359317. Epub 2017 Sep 14.
In the present study, the in vitro anticancer (antiproliferative) effects of Bacillus coagulans Unique IS2 were evaluated on human colon cancer (COLO 205), cervical cancer (HeLa), and chronic myeloid leukemia (K562) cell lines with a human embryonic kidney cell line (HEK 293T) as noncancerous control cells. The Cytotoxicity assay (MTT) clearly demonstrated a 22%, 31.7%, and 19.5% decrease in cell proliferation of COLO 205, HeLa, and K562 cells, respectively, when compared to the noncancerous HEK 293T cells. Normal phase-contrast microscopic images clearly suggested that the mechanism of cell death is by apoptosis. To further confirm the induction of apoptosis by Unique IS2, the sub-G0-G1 peak of the cell cycle was quantified using a flow cytometer and the data indicated 40% of the apoptotic cells in Unique IS2-treated COLO cells when compared with their untreated control cells. The Western blot analysis showed an increase in pro-apoptotic protein BAX, decrease in antiapoptotic protein, Bcl2, decrease in mitochondrial membrane potential, increase in cytochrome c release, increase in Caspase 3 activity, and cleavage of poly(ADP-ribose) polymerase. The present study suggests that the heat-killed culture supernatant of B. coagulans can be more effective in inducing apoptosis of colon cancer cells and that can be considered for adjuvant therapy in the treatment of colon carcinoma.
在本研究中,以人胚肾细胞系(HEK 293T)作为非癌细胞对照,评估了凝结芽孢杆菌Unique IS2对人结肠癌细胞系(COLO 205)、宫颈癌细胞系(HeLa)和慢性髓性白血病细胞系(K562)的体外抗癌(抗增殖)作用。细胞毒性测定(MTT)清楚地表明,与非癌性HEK 293T细胞相比,COLO 205、HeLa和K562细胞的细胞增殖分别降低了22%、31.7%和19.5%。正常相差显微镜图像清楚地表明细胞死亡机制是凋亡。为了进一步证实Unique IS2诱导凋亡,使用流式细胞仪对细胞周期的亚G0-G1峰进行定量,数据表明与未处理的对照细胞相比,Unique IS2处理的COLO细胞中有40%的凋亡细胞。蛋白质免疫印迹分析显示促凋亡蛋白BAX增加、抗凋亡蛋白Bcl2减少、线粒体膜电位降低、细胞色素c释放增加、半胱天冬酶3活性增加以及聚(ADP-核糖)聚合酶裂解。本研究表明,凝结芽孢杆菌的热灭活培养上清液在诱导结肠癌细胞凋亡方面可能更有效,可考虑用于结肠癌治疗的辅助治疗。