Betulinic acid inhibits stemness and EMT of pancreatic cancer cells via activation of AMPK signaling

Sun,Liankang; Cao,Junyu; Chen,Ke; Cheng,Liang; Zhou,Cancan; Yan,Bin; Qian,Weikun; Li,Jie; Duan,Wanxing; Ma,Jiguang; Qi,Dan; Wu,Erxi; Wang,Zheng; Liu,Qingguang; Ma,Qingyong; Xu,Qinhong

doi:10.3892/ijo.2018.4604

Betulinic acid inhibits stemness and EMT of pancreatic cancer cells via activation of AMPK signaling

Authors:
- Liankang Sun
- Junyu Cao
- Ke Chen
- Liang Cheng
- Cancan Zhou
- Bin Yan
- Weikun Qian
- Jie Li
- Wanxing Duan
- Jiguang Ma
- Dan Qi
- Erxi Wu
- Zheng Wang
- Qingguang Liu
- Qingyong Ma
- Qinhong Xu
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Affiliations: Department of Hepatobiliary Surgery, First Affiliated Hospital of Xi'an Jiaotong University, Xi'an, Shaanxi 710061, P.R. China, Department of Anesthesiology, First Affiliated Hospital of Xi'an Jiaotong University, Xi'an, Shaanxi 710061, P.R. China, 3Department of Neurosurgery, Baylor Scott & White Health, Temple, TX 78508, USA, Department of Geriatric Surgery, First Affiliated Hospital of Xi'an Jiaotong University, Xi'an, Shaanxi 710061, P.R. China
Published online on: October 24, 2018 https://doi.org/10.3892/ijo.2018.4604
Pages: 98-110
Copyright: © Sun et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Cancer stem cells (CSCs), which are found in various types of human cancer, including pancreatic cancer, possess elevated metastatic potential, lead to tumor recurrence and cause chemoradiotherapy resistance. Alterations in cellular bioenergetics through the regulation of 5' adenosine monophosphate‑activated protein kinase (AMPK) signaling may be a prerequisite to stemness. Betulinic acid (BA) is a well‑known bioactive compound with antiretroviral and anti‑inflammatory potential, which has been reported to exert anticancer effects on various types of cancer, including pancreatic cancer. The present study aimed to investigate whether BA could inhibit pancreatic CSCs via regulation of AMPK signaling. The proliferation of pancreatic cancer cells was examined by MTT and colony formation assays. The migratory and invasive abilities of pancreatic cancer cells were assessed using wound‑scratch and Transwell invasion assays. In addition, the expression levels of candidate genes were measured by reverse transcription‑quantitative polymerase chain reaction and western blotting. The results revealed that BA inhibited the proliferation and tumorsphere formation of pancreatic cancer cells, suppressed epithelial‑mesenchymal transition (EMT), migration and invasion, and reduced the expression of three pluripotency factors [SRY‑box 2 (Sox2), octamer‑binding protein 4 (Oct4) and Nanog]. Furthermore, immunohistochemical analysis confirmed that there was a significant inverse association between the expression levels of phosphorylated (P)‑AMPK and Sox2 in pancreatic cancer, and it was revealed that BA may activate AMPK signaling. Notably, knockdown of AMPK reversed the suppressive effects of BA on EMT and stemness of pancreatic cancer cells. In addition, BA reversed the effects of gemcitabine on stemness and enhanced the sensitivity of pancreatic cancer cells to gemcitabine. Collectively, these results indicated that BA may effectively inhibit pluripotency factor expression (Sox2, Oct4 and Nanog), EMT and the stem‑like phenotype of pancreatic cancer cells via activating AMPK signaling. Therefore, BA may be considered an attractive therapeutic candidate and an effective inhibitor of the stem‑like phenotype in pancreatic cancer cells. Further investigation into the development of BA as an anticancer drug is warranted.

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