Abstract：OBJECTIVE To investigate the effect of bufalin on the proliferation of esophageal cancer and its possible molecular mechanism. METHODS The effects of bufalin on the activity of esophageal cancer cell line KYSE-70 were determined by MTT assay and LDH assay kit. Colony-forming and EdU (5-ethynyl-2-deoxyuridine) assay were performed to detect the inhibitory effect of bufalin on the proliferation of KYSE-70 cells. DAPI staining, TUNEL and flow cytometry were used to assess the effects of bufalin on the apoptosis of KYSE-70 cells. Quantitative real-time PCR (qPCR) and Western blotting were used to determine the relevant expression changes in mRNA and protein of apoptosis-related factors Bcl-2, Bax, NF-κBp65, NF-κBpp65. The mitochondrial function changes of KYSE-70 cells were studied by using JC-1 fluorescent probe kit and ATP detection kit after bufalin treatment. Finally, the effect of bufalin on esophageal cancer proliferation in vivo was studied by xenograft model in nude mice. RESULTS The results of MTT and LDH assay shown that bufalin inhibited the activity of KYSE-70 cells. Colony-forming and EdU assay showed that bufalin significantly suppressed KYSE-70 cells proliferation. DAPI staining showed that chromatin heterogeneity, nuclear concentration and fragmentation were observed after bufalin treatment. It was found that bufalin treatment significantly promoted KYSE-70 cells apoptosis by TUNEL staining and flow cytometry assay of qPCR and Western blot showed that Bcl-2 expression was down-regulated, Bax expression was up-regulated and Bax/ Bcl-2 expression ratio was increased after bufalin treatment. The mitochondrial membrane potential and the ATP production of KYSE-70 cells were significant reduced after bufalin treatment. In vivo, the growth of xenograft tumors was significantly inhibited in bufalin group. CONCLUSION Bufalin markedly inhibits KYSE-70 cells proliferation by promoting apoptosis, and the possible mechanism of apoptosis may be related to mitochondrial pathway. Our results indicate that bufalin may be a potential therapeutic agent for esophageal cancer.
陈慧, 聂雪珂, 杨雅亭, 吴远远, 王佳鑫, 牛梦兰, 刘红亮, 鲍登克. 蟾毒灵对食管癌增殖的影响及机制研究[J]. 中国药学杂志, 2020, 55(16): 1346-1353.
CHEN Hui, NIE Xue-ke, YANG Ya-ting, WU Yuan-yuan, WANG Jia-xin, NIU Meng-lan, LIU Hong-liang, BAO Deng-ke. Effects of Bufalin on Human Esophageal Cancer Cells Proliferation and Mechanism Study. Chinese Pharmaceutical Journal, 2020, 55(16): 1346-1353.
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2020, Vol. 55 
