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

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

Research output: Contribution to journalArticle

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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.

Original languageEnglish (US)
Pages (from-to)98-110
Number of pages13
JournalInternational journal of oncology
Volume54
Issue number1
DOIs
StatePublished - Jan 2019

Fingerprint

Epithelial-Mesenchymal Transition
Adenosine Monophosphate
Pancreatic Neoplasms
Protein Kinases
gemcitabine
Octamer Transcription Factor-3
Neoplastic Stem Cells
betulinic acid
Phenotype
Neoplasms
Chemoradiotherapy
Energy Metabolism
Reverse Transcription
Anti-Inflammatory Agents
Western Blotting

Keywords

  • 5' adenosine monophosphate-activated protein kinase
  • Betulinic acid
  • Cancer stem cells
  • Epithelial-mesenchymal transition
  • Gemcitabine
  • Pancreatic cancer

ASJC Scopus subject areas

  • Oncology
  • Cancer Research

Cite this

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.

In: International journal of oncology, Vol. 54, No. 1, 01.2019, p. 98-110.

Research output: Contribution to journalArticle

Sun, L, Cao, J, Chen, K, Cheng, L, Zhou, C, Yan, B, Qian, W, Li, J, Duan, W, Ma, J, Qi, D, Wu, E, Wang, Z, Liu, Q, Ma, Q & Xu, Q 2019, 'Betulinic acid inhibits stemness and EMT of pancreatic cancer cells via activation of AMPK signaling', International journal of oncology, vol. 54, no. 1, pp. 98-110. https://doi.org/10.3892/ijo.2018.4604
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. / Betulinic acid inhibits stemness and EMT of pancreatic cancer cells via activation of AMPK signaling. In: International journal of oncology. 2019 ; Vol. 54, No. 1. pp. 98-110.
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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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AU - Yan, Bin

AU - Qian, Weikun

AU - Li, Jie

AU - Duan, Wanxing

AU - Ma, Jiguang

AU - Qi, Dan

AU - Wu, Erxi

AU - Wang, Zheng

AU - Liu, Qingguang

AU - Ma, Qingyong

AU - Xu, Qinhong

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