Guang-Yi Gao1, Jun Ma2, Peng Lu3, Xuan Jiang4, Cheng Chang5. 1. Department of Traditional Chinese Medicine, Huai'an Second People's Hospital, The Affiliated Huai'an Hospital of Xuzhou Medical University, No. 62, Huaihai South Road, 223002, Huai'an, Jiangsu, China. Electronic address: guangyi_gg@sina.com. 2. Department of Oncology, Huai'an Hospital of Chinese Medicine, The Affiliated Huai'an Hospital of Nanjing University of Chinese Medicine, Nanjing, Jiangsu, China. 3. Department of Pharmacy, Huai'an Maternity and Child Healthcare Hospital Affiliated to Yangzhou University Medical Academy, Huai'an, Jiangsu, China. 4. Department of Oncology, Huai'an Second People's Hospital, The Affiliated Huai'an Hospital of Xuzhou Medical University, Huai'an, Jiangsu, China. 5. Internal Medicine of Traditional Chinese Medicine, Nanjing Jianzhong Hospital of Traditional Chinese Medicine, Nanjing, Jiangsu, China.
Abstract
OBJECTIVE: To investigate the effect of Ophiopogonin B (OP-B) on the autophagy and apoptosis of colon cancer cells via the regulation of JNK/c-Jun signaling pathway. METHODS: Colon cancer cell lines (HT-29 and HCT-116) were treated with various concentrations of OP-B (0, 5, 10and 20 μmol/l) and JNK inhibitor SP600125. MTT assay, flow cytometry, immunofluorescence staining were used to detect the biological function ofHT-29 and HCT-116 cells, and expressions of autophagy-,apoptotic- and pathway-related proteins were measured by Western Blot. Moreover, a nude mice model with transplanted tumor was used to observe the effect of OP-B on the growth, autophagy and apoptosis of the transplanted tumor of colon cancer. RESULTS: The results demonstrated that OP-B suppressed the proliferation of HT-29 and HCT-116 cell lines through the G0/G1 phase cell cycle arrest. Moreover, OP-B induced apoptosis by inhibiting the expression of Bax and cleaved caspase 3 and promoting the expression of Bcl-2. Treatment with OP-B also increased the expression of Beclin 1 and the conversion of LC3I to LC3II with the activation of JNK/c-Jun signaling pathway, but reduced the expression of P62, whereas SP600125 (an inhibitor of JNK) reversed these process. In addition, the xenograft model using HCT-116 cells provided further evidence of the inhibition of OP-B on tumor proliferation. Immunohistochemistry detection verified that OP-B enhance the positive expression rate of LC3, and increase the apoptosis index of tumor cells in vivo. Importantly, all these changes induced by OP-B were clearly in a dose-dependent manner. CONCLUSION: OP-B may induce cell autophagy, apoptosis and cell cycle arrest by activating the JNK/ c-Jun signaling pathway, thereby inhibiting the growth of colon cancer.
OBJECTIVE: To investigate the effect of Ophiopogonin B (OP-B) on the autophagy and apoptosis of colon cancer cells via the regulation of JNK/c-Jun signaling pathway. METHODS: Colon cancer cell lines (HT-29 and HCT-116) were treated with various concentrations of OP-B (0, 5, 10and 20 μmol/l) and JNK inhibitor SP600125. MTT assay, flow cytometry, immunofluorescence staining were used to detect the biological function ofHT-29 and HCT-116 cells, and expressions of autophagy-,apoptotic- and pathway-related proteins were measured by Western Blot. Moreover, a nude mice model with transplanted tumor was used to observe the effect of OP-B on the growth, autophagy and apoptosis of the transplanted tumor of colon cancer. RESULTS: The results demonstrated that OP-B suppressed the proliferation of HT-29 and HCT-116 cell lines through the G0/G1 phase cell cycle arrest. Moreover, OP-B induced apoptosis by inhibiting the expression of Bax and cleaved caspase 3 and promoting the expression of Bcl-2. Treatment with OP-B also increased the expression of Beclin 1 and the conversion of LC3I to LC3II with the activation of JNK/c-Jun signaling pathway, but reduced the expression of P62, whereas SP600125 (an inhibitor of JNK) reversed these process. In addition, the xenograft model using HCT-116 cells provided further evidence of the inhibition of OP-B on tumor proliferation. Immunohistochemistry detection verified that OP-B enhance the positive expression rate of LC3, and increase the apoptosis index of tumor cells in vivo. Importantly, all these changes induced by OP-B were clearly in a dose-dependent manner. CONCLUSION: OP-B may induce cell autophagy, apoptosis and cell cycle arrest by activating the JNK/ c-Jun signaling pathway, thereby inhibiting the growth of colon cancer.
Authors: Jun Qian; Yi Cao; Junfeng Zhang; Lingchang Li; Juan Wu; Guoli Wei; Jialin Yu; Jiege Huo Journal: Transl Cancer Res Date: 2020-11 Impact factor: 1.241