Codonopis bulleynana Forest ex Diels inhibits autophagy and induces apoptosis of colon cancer cells by activating the NF-κB signaling pathway

Luan,Yunpeng; Li,Yanmei; Zhu,Lina; Zheng,Shuangqing; Mao,Dechang; Chen,Zhuxue; Cao,Yong

doi:10.3892/ijmm.2017.3337

Codonopis bulleynana Forest ex Diels inhibits autophagy and induces apoptosis of colon cancer cells by activating the NF-κB signaling pathway

Authors:
- Yunpeng Luan
- Yanmei Li
- Lina Zhu
- Shuangqing Zheng
- Dechang Mao
- Zhuxue Chen
- Yong Cao
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Affiliations: Department of Life Technology Teaching and Research, School of Life Science, Southwest Forestry University, Kunming, Yunnan 652400, P.R. China, Kunming Pharmaceutical Corp., Kunming, Yunnan 652400, P.R. China
Published online on: December 21, 2017 https://doi.org/10.3892/ijmm.2017.3337
Pages: 1305-1314
Copyright: © Luan et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Despite its favorable clinical efficacy, oxaliplatin‑based chemotherapy frequently results in treatment withdrawal and induces liver damage in colon cancer. Therefore, it is important to develop novel drugs, which can safely and effectively complement or replace the therapeutic effects of oxaliplatin. Codonopis bulleynana Forest ex Diels (cbFeD) has wide range of pharmacological effects, including anticancer effects. In the present study, the anticancer activity of cbFeD and its potential molecular mechanisms were investigated. In vitro, cell counting kit‑8 assays and flow cytometry were used to assess the anti‑proliferation and apoptosis‑promoting activities of cbFeD. Transmission electron microscopy was used to monitor the autophagic vesicles. Immunofluorescence staining was performed to observe the nuclear translocation of p65 and the fluorescence of microtubule‑associated protein 1 light chain 3 (LC3) B‑II. The protein expression levels of p65, inhibitor of nuclear factor (NF)‑κB (IκB) a, LC3B‑I, LC3B‑II and Beclin‑1 were detected using western blot analysis. In vivo, the antitumor effect of cbFeD was assessed in colon cancer‑bearing nude mice as a model. H&E staining and immunohistochemistry (IHC) were performed, with oxaliplatin set as a positive control. The results showed that cbFeD inhibited cell proliferation and promoted cell apoptosis in a dose‑dependent manner. The effects of a high dose of cbFeD on colon cancer cells were similar to those of oxaliplatin. In HCT116 and SW480 cells, cbFeD inhibited the expression of IκBα, LC3B‑I/II and Beclin‑1, and the results of western blot analysis and immunofluorescence showed that, in the cells treated with cbFeD, p65 gradually entered nuclei in a dose‑dependent manner, and the expression of LC3B‑II was gradually reduced. The results of the acridine orangestaining and electron microscopy demonstrated fewer autophagic vesicles in the high‑dose cbFeD group and the oxaliplatin group. The high dose of cbFeD reversed the effect of pyrrolidine dithiocarbamate, a p65‑inhibitor, on the expression of p65, LC3B‑I, LC3B‑II and Beclin‑1, and on the production of autophagic vacuoles. The high dose of cbFeD and oxaliplatin also suppressed tumorigenicity in vivo. The results of the H&E and IHC staining confirmed the inhibition of autophagy (LC3 and Beclin‑1) and activation of p65 by treatment with the high dose of cbFeD and oxaliplatin. Taken together, cbFeD exhibited an antitumor effect in colon cancer cells by inhibiting autophagy through activation of the NF‑κB pathway. Therefore, cbFeD may be a promising Chinese herbal compound for development for use in cancer therapy.

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1	Arber N and Levin B: Chemoprevention of colorectal neoplasia: The potential for personalized medicine. Gastroenterology. 134:1224–1237. 2008. View Article : Google Scholar : PubMed/NCBI
2	Yang J, Du XL, Li ST, Wang BY, Wu YY, Chen ZL, Lv M, Shen YW, Wang X, Dong DF, et al: Characteristics of differently located colorectal cancers support proximal and distal classification: A population-based study of 57,847 patients. PLoS One. 11:e01675402016. View Article : Google Scholar : PubMed/NCBI
3	Katz ML, Young GS, Zimmermann BJ, Tatum CM and Paskett ED: Assessing colorectal cancer screening barriers by two methods. J Cancer Educ. Dec 8–2016.Epub ahead of print. View Article : Google Scholar : PubMed/NCBI
4	Alajez NM: Large-scale analysis of gene expression data reveals a novel gene expression signature associated with colorectal cancer distant recurrence. PLoS One. 11:e01674552016. View Article : Google Scholar : PubMed/NCBI
5	Rocha R, Marinho R, Aparicio D, Fragoso M, Sousa M, Gomes A, Leichsenring C, Carneiro C, Geraldes V and Nunes V: Impact of bowel resection margins in node negative colon cancer. Springerplus. 5:19592016. View Article : Google Scholar : PubMed/NCBI
6	Panteleimonitis S, Ahmed J, Harper M and Parvaiz A: Critical analysis of the literature investigating urogenital function preservation following robotic rectal cancer surgery. World J Gastrointest Surg. 8:744–754. 2016. View Article : Google Scholar : PubMed/NCBI
7	Sommerer C and Zeier M: Clinical manifestation and management of ADPKD in Western countries. Kidney Dis (Basel). 2:120–127. 2016. View Article : Google Scholar
8	Liu D and Liang XC: New developments in the pharmacodynamics and pharmacokinetics of combination of Chinese medicine and Western medicine. Chin J Integr Med. 23:312–319. 2017. View Article : Google Scholar
9	Pinhua L, Yarong J, Mingyan L, Shirui L and Yaguan Z: Total flavonoids and antioxidant activity of aerial parts of Codonopsis bulleyana. Southwest China J Agricultural Sci. 27:1894–1898. 2014.
10	Chen Z, Li Y, Lu L, Lu B, Zhou J and Hu Y: Pharmacodynamic study on Qi-Blood-Enriching effects of Codonopis bulleynana Fores tex Diels. Shanghai Zhong Yi Xue Yuan. 43:68–71. 2009.
11	Chen Z, Li Y, Zhou J, Wang Y, Lu B, Lu Z and Yuan J: Chou can bu tong bu wei ti qu wu dui qi xu yu xue xu dong wu mo xing de ying xiang. J Chin Med Materials. 11:1731–1733. 2009.In Chinese.
12	Dong LD, Qian ZG, Yang Z and Cheng YX: Study on the anti-hyperlipidemia, anti-fatigue and anti-anoxia effects of Codonopsis foetens Hook. Yunnan Chin Med J. 36:66–68. 2015.In Chinese.
13	Chen ZJ, Li YS, Wei QH, Chen SL and Chen DX: Effects of Codonopis bulleynana Forest ex Diels on enhancing sensitivity, reducing toxicity of chemotherapy and regulating immune function in Sarcoma 180 tumor-bearing mice. Chin Traditional Patent Med. 34:1848–1851. 2012.In Chinese.
14	Whitehurst C, Pantelides ML, Moore JV, Brooman PJ and Blacklock NJ: In vivo laser light distribution in human prostatic carcinoma. J Urol. 151:1411–1415. 1994. View Article : Google Scholar : PubMed/NCBI
15	Xiong L, Liu Z, Ouyang G, Lin L, Huang H, Kang H, Chen W, Miao X and Wen Y: Autophagy inhibition enhances photocytotoxicity of Photosan-II in human colorectal cancer cells. Oncotarget. 8:6419–6432. 2017.
16	Shintani T and Klionsky DJ: Autophagy in health and disease: A double-edged sword. Science. 306:990–995. 2004. View Article : Google Scholar : PubMed/NCBI
17	Rubinsztein DC, Gestwicki JE, Murphy LO and Klionsky DJ: Potential therapeutic applications of autophagy. Nat Rev Drug Discov. 6:304–312. 2007. View Article : Google Scholar : PubMed/NCBI
18	Panganiban RA, Snow AL and Day RM: Mechanisms of radiation toxicity in transformed and non-transformed cells. Int J Mol Sci. 14:15931–15958. 2013. View Article : Google Scholar : PubMed/NCBI
19	Chen ST, Lee TY, Tsai TH, Lin YC, Lin CP, Shieh HR, Hsu ML, Chi CW, Lee MC, Chang HH and Chen YJ: The Traditional Chinese Medicine DangguiBuxue tang sensitizes colorectal cancer cells to chemoradiotherapy. Molecules. 21:pii:E1677. 2016. View Article : Google Scholar
20	Wang M, Tan W, Zhou J, Leow J, Go M, Lee HS and Casey PJ: A small molecule inhibitor of isoprenylcysteine carboxymethyltransferase induces autophagic cell death in PC3 prostate cancer cells. J Biol Chem. 283:18678–18684. 2008. View Article : Google Scholar : PubMed/NCBI
21	Wang B, Qiao L, Shi Y, Feng X, Chen D and Guo H: ASPP2 inhibits oxaliplatin-induced autophagy and promotes apoptosis of colon cancer cells. Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 31:898–904. 2015.In Chinese. PubMed/NCBI
22	Tan S, Peng X, Peng W, Zhao Y and Wei Y: Enhancement of oxaliplatin-induced cell apoptosis and tumor suppression by 3-methyladenine in colon cancer. Oncol Lett. 9:2056–2062. 2015. View Article : Google Scholar : PubMed/NCBI
23	O’Donovan TR, Rajendran S, O’Reilly S, O’Sullivan GC and McKenna SL: Lithium modulates autophagy in esophageal and colorectal cancer cells and enhances the efficacy of therapeutic agents in vitro and in vivo. PLoS One. 10:e01346762015. View Article : Google Scholar
24	Shan J, Xuan Y, Zhang Q, Zhu C, Liu Z and Zhang S: Ursolic acid synergistically enhances the therapeutic effects of oxaliplatin in colorectal cancer. Protein Cell. 7:571–585. 2016. View Article : Google Scholar : PubMed/NCBI
25	Shan J, Xuan Y, Zhang Q, Zhu C, Liu Z, Zhang S, et al: Inhibition of the NF-κB pathway by nafamostat mesilate suppresses colorectal cancer growth and metastasis. Cancer Lett. 380:87–97. 2016. View Article : Google Scholar
26	Zeng Y, Liu XH, Tarbell J and Fu B: Sphingosine 1-phosphate induced synthesis of glycocalyx on endothelial cells. Exp Cell Res. 339:90–95. 2015. View Article : Google Scholar : PubMed/NCBI
27	Paglin S, Hollister T, Delohery T, Hackett N, McMahill M, Sphicas E, Domingo D and Yahalom J: A novel response of cancer cells to radiation involves autophagy and formation of acidic vesicles. Cancer Res. 61:439–444. 2001.PubMed/NCBI
28	Dutta D, Chakraborty B, Sarkar A, Chowdhury C and Das P: A potent betulinic acid analogue ascertains an antagonistic mechanism between autophagy and proteasomal degradation pathway in HT-29 cells. BMC Cancer. 16:232016. View Article : Google Scholar : PubMed/NCBI
29	Chen SL, Cai SR, Zhang XH, Li WF, Zhai ET, Peng JJ, Wu H, Chen CQ, Ma JP, Wang Z and He YL: Targeting CRMP-4 by lentivirus-mediated RNA interference inhibits SW480 cell proliferation and colorectal cancer growth. Exp Ther Med. 12:2003–2008. 2016. View Article : Google Scholar : PubMed/NCBI
30	Jiang H, Ju H, Zhang L, Lu H and Jie K: microRNA-577 suppresses tumor growth and enhances chemosensitivity in colorectal cancer. J Biochem Mol Toxicol. Feb 2–2017.Epub ahead of print. View Article : Google Scholar
31	Baldwin AS: Regulation of cell death and autophagy by IKK and NF-kappaB: Critical mechanisms in immune function and cancer. Immunol Rev. 246:327–345. 2012. View Article : Google Scholar : PubMed/NCBI
32	Samuel T, Fadlalla K, Gales DN, Putcha BD and Manne U: Variable NF-κB pathway responses in colon cancer cells treated with chemotherapeutic drugs. BMC Cancer. 14:5992014. View Article : Google Scholar