Asiatic acid inhibits proliferation, migration and induces apoptosis by regulating Pdcd4 via the PI3K/Akt/mTOR/p70S6K signaling pathway in human colon carcinoma cells

Hao,Yajuan; Huang,Jiawei; Ma,Yun; Chen,Wancheng; Fan,Qin; Sun,Xuegang; Shao,Meng; Cai,Hongbing

doi:10.3892/ol.2018.8417

Asiatic acid inhibits proliferation, migration and induces apoptosis by regulating Pdcd4 via the PI3K/Akt/mTOR/p70S6K signaling pathway in human colon carcinoma cells

Authors:
- Yajuan Hao
- Jiawei Huang
- Yun Ma
- Wancheng Chen
- Qin Fan
- Xuegang Sun
- Meng Shao
- Hongbing Cai
View Affiliations

Affiliations: Department of Traditional Chinese Medicine, Southern Medical University, Guangzhou, Guangdong 510515, P.R. China, Department of Pharmacy, Nanfang Hospital of Southern Medical University, Guangzhou, Guangdong 510515, P.R. China, Cancer Research Institute, Southern Medical University, Guangzhou, Guangdong 510515, P.R. China
Published online on: April 4, 2018 https://doi.org/10.3892/ol.2018.8417
Pages: 8223-8230

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Previous studies have demonstrated that asiatic acid (AA), the major component of Centella asiatica, is able to meditate cytotoxic and anticancer effects on various types of carcinoma cells. In order to investigate the molecular mechanism that underlies the antitumor effect of AA, the present study investigated the effects of AA on proliferation, migration and apoptosis of SW480 and HCT116 colon cancer cells. Viability and changes in cell morphology in the cells were assessed by MTT assay and transmission electron microscopy, respectively. Colony formation analysis was used to observe proliferation of the single cell, and migratory ability of the cells was assessed by performing Transwell migration assay. Hoechst 33342 nuclear staining and flow cytometry were used to assess apoptosis in colon carcinoma cells. The expression of proteins associated with the phosphoinositide 3‑kinase (PI3K)/protein kinase B (Akt)/mammalian target of rapamycin (mTOR)/p70S6K signaling pathway and epithelial‑mesenchymal transition (EMT) marker were analyzed by western blotting. The present study revealed that proliferation and migration of colon carcinoma cells were inhibited by AA in a dose‑dependent and time‑dependent manner. Numerous apoptotic bodies were observed, and G2/M and S phase progression were delayed in colon cancer cells treated with AA, but not in the control group. A number of phosphorylated proteins, including PI3K, Akt (Ser473), mTOR, ribosomal protein S6 kinase (p70S6K) downregulated, while the expression of Pdcd4 was upregulated following treatment with AA. Additionally, AA affects expression of EMT markers in a dose‑dependent manner. On the basis of these results, it was concluded that AA inhibited proliferation, migration and induced apoptosis of colon cancer cells by regulating Pdcd4 via the PI3K/Akt/mTOR/p70S6K signaling pathway. These observations suggest that AA may be a potential therapeutic agent for the treatment of colon carcinoma.

View Figures

View References

1	Brenner H, Kloor M and Pox CP: Colorectal cancer. Lancet. 383:1490–1502. 2014. View Article : Google Scholar : PubMed/NCBI
2	Du C, Huang D, Peng Y, Yao Y, Zhao Y, Yang Y, Wang H, Cao L, Zhu WG and Gu J: 5-Fluorouracil targets histone acetyltransferases p300/CBP in the treatment of colorectal cancer. Cancer Lett. 400:183–193. 2017. View Article : Google Scholar : PubMed/NCBI
3	Woo IS and Jung YH: Metronomic chemotherapy in metastatic colorectal cancer. Cancer Lett. 400:319–324. 2017. View Article : Google Scholar : PubMed/NCBI
4	Siegel R, Desantis C and Jemal A: Colorectal cancer statistics, 2014. CA Cancer J Clin. 64:104–117. 2014. View Article : Google Scholar : PubMed/NCBI
5	Weidle UH, Birzele F and Krüger A: Molecular targets and pathways involved in liver metastasis of colorectal cancer. Clin Exp Metastasis. 32:623–635. 2015. View Article : Google Scholar : PubMed/NCBI
6	Nasir MN, Habsah M, Zamzuri I, Rammes G, Hasnan J and Abdullah J: Effects of asiatic acid on passive and active avoidance task in male spraque-dawley rats. J Ethnopharmacol. 134:203–209. 2011. View Article : Google Scholar : PubMed/NCBI
7	Zhang X, Wu J, Dou Y, Xia B, Rong W, Rimbach G and Lou Y: Asiatic acid protects primary neurons against C2-ceramide-induced apoptosis. Eur J Pharmacol. 679:51–59. 2012. View Article : Google Scholar : PubMed/NCBI
8	Lee Soo Y, Jin DQ, Beak SM, Lee ES and Kim JA: Inhibition of ultraviolet-A-modulated signaling pathways by asiatic acid and ursolic acid in HaCaT human keratinocytes. Eur J Pharmacol. 476:173–178. 2003. View Article : Google Scholar : PubMed/NCBI
9	Lee YS, Jin DQ, Kwon EJ, Park SH, Lee ES, Jeong TC, Nam DH, Huh K and Kim JA: Asiatic acid, a triterpene, induces apoptosis through intracellular Ca2⁺ release and enhanced expression of p53 in HepG2 human hepatoma cells. Cancer Lett. 186:83–91. 2002. View Article : Google Scholar : PubMed/NCBI
10	Hsu YL, Kuo PL, Lin LT and Lin CC: Asiatic acid, a triterpene, induces apoptosis and cell cycle arrest through activation of extracellular signal-regulated kinase and p38 mitogen-activated protein kinase pathways in human breast cancer cells. J Pharmacol Exp Ther. 313:333–344. 2005. View Article : Google Scholar : PubMed/NCBI
11	Adtani PN, Narasimhan M, Punnoose AM and Kambalachenu HR: Antifibrotic effect of Centella asiatica Linn and asiatic acid on arecoline-induced fibrosis in human buccal fibroblasts. J Investig Clin Dent. 8:2017. View Article : Google Scholar : PubMed/NCBI
12	Wang L, Xu J, Zhao C, Zhao L and Feng B: Antiproliferative, cell-cycle dysregulation effects of novel asiatic acid derivatives on human non small cell lung cancer cells. Chem Pharm Bull (Tokyo). 61:1015–1023. 2013. View Article : Google Scholar : PubMed/NCBI
13	Gurfinkel DM, Chow S, Hurren R, Gronda M, Henderson C, Berube C, Hedley DW and Schimmer AD: Disruption of the endoplasmic reticulum and increases in cytoplasmic calcium are early events in cell death induced by the natural triterpenoid Asiatic acid. Apoptosis. 11:1463–1471. 2006. View Article : Google Scholar : PubMed/NCBI
14	Thakor FK, Wan KW, Welsby PJ and Welsby G: Pharmacological effects of asiatic acid in glioblastoma cells under hypoxia. Mol Cell Biochem. 430:179–190. 2017. View Article : Google Scholar : PubMed/NCBI
15	Kavitha CV, Jain AK, Agarwal C, Pierce A, Keating A, Huber KM, Serkova NJ, Wempe MF, Agarwal R and Deep G: Asiatic acid induces endoplasmic reticulum stress and apoptotic death in glioblastoma multiforme cells both in vitro and in vivo. Mol Carcinog. 54:1417–1429. 2015. View Article : Google Scholar : PubMed/NCBI
16	Park BC, Bosire KO, Lee ES, Lee YS and Kim JA: Asiatic acid induces apoptosis in SK-MEL-2 human melanoma cells. Cancer Lett. 218:81–90. 2005. View Article : Google Scholar : PubMed/NCBI
17	Wu Q, Lv T, Chen Y, Wen L, Zhang J, Jiang X and Liu F: Apoptosis of HL-60 human leukemia cells induced by Asiatic acid through modulation of B-cell lymphoma 2 family proteins and the mitogen-activated protein kinase signaling pathway. Mol Med Rep. 12:1429–1434. 2015. View Article : Google Scholar : PubMed/NCBI
18	Xu MF, Xiong YY, Liu JK, Qian JJ, Zhu L and Gao J: Asiatic acid, a pentacyclic triterpene in Centella asiatica, attenuates glutamate-induced cognitive deficits in mice and apoptosis in SH-SY5Y cells. Acta Pharmacol Sin. 33:578–587. 2012. View Article : Google Scholar : PubMed/NCBI
19	Tang XL, Yang XY, Jung HJ, Kim SY, Jung SY, Choi DY, Park WC and Park H: Asiatic acid induces colon cancer cell growth inhibition and apoptosis through mitochondrial death cascade. Biol Pharm Bull. 32:1399–1405. 2009. View Article : Google Scholar : PubMed/NCBI
20	Kiesslich T, Pichler M and Neureiter D: Epigenetic control of epithelial-mesenchymal-transition in human cancer. Mol Clin Oncol. 1:3–11. 2013. View Article : Google Scholar : PubMed/NCBI
21	Gheldof A and Berx G: Cadherins and epithelial-to-mesenchymal transition. Prog Mol Biol Transl Sci. 116:317–336. 2013. View Article : Google Scholar : PubMed/NCBI
22	Bao B, Wang Z, Ali S, Kong D, Li Y, Ahmad A, Banerjee S, Azmi AS, Miele L and Sarkar FH: Notch-1 induces epithelial-mesenchymal transition consistent with cancer stem cell phenotype in pancreatic cancer cells. Cancer Lett. 307:26–36. 2011. View Article : Google Scholar : PubMed/NCBI
23	Zavadil J, Narasimhan M, Blumenberg M and Schneider RJ: Transforming growth factor-beta and microRNA: mRNA regulatory networks in epithelial plasticity. Cells Tissues Organs. 185:157–161. 2007. View Article : Google Scholar : PubMed/NCBI
24	Syed DN, Afaq F, Sarfaraz S, Khan N, Kedlaya R, Setaluri V and Mukhtar H: Delphinidin inhibits cell proliferation and invasion via modulation of Met receptor phosphorylation. Toxicol Appl Pharmacol. 231:52–60. 2008. View Article : Google Scholar : PubMed/NCBI
25	Ren W, Liu Y, Wan S, Fei C, Wang W, Chen Y, Zhang Z, Wang T, Wang J, Zhou L, et al: BMP9 inhibits proliferation and metastasis of HER2-positive SK-BR-3 breast cancer cells through ERK1/2 and PI3K/AKT pathways. PLoS One. 9:e968162014. View Article : Google Scholar : PubMed/NCBI
26	Wang H, Duan L, Zou Z, Li H, Yuan S, Chen X, Zhang Y, Li X, Sun H, Zha H, et al: Activation of the PI3K/Akt/mTOR/p70S6K pathway is involved in S100A4-induced viability and migration in colorectal cancer cells. Int J Med Sci. 11:841–849. 2014. View Article : Google Scholar : PubMed/NCBI
27	Wen YH, Shi X, Chiriboga L, Matsahashi S, Yee H and Afonja O: Alterations in the expression of PDCD4 in ductal carcinoma of the breast. Oncol Rep. 18:1387–1393. 2007.PubMed/NCBI
28	Afonja O, Juste D, Das S, Matsuhashi S and Samuels HH: Induction of PDCD4 tumor suppressor gene expression by RAR agonists, antiestrogen and HER-2/neu antagonist in breast cancer cells. Evidence for a role in apoptosis. Oncogene. 23:8135–8145. 2004. View Article : Google Scholar : PubMed/NCBI
29	Leupold JH, Yang HS, Colburn NH, Asangani I, Post S and Allgayer H: Tumor suppressor Pdcd4 inhibits invasion/intravasation and regulates urokinase receptor (u-PAR) gene expression via Sp-transcription factors. Oncogene. 26:4550–4562. 2007. View Article : Google Scholar : PubMed/NCBI
30	Yang HS, Jansen AP, Nair R, Shibahara K, Verma AK, Cmarik JL and Colburn NH: A novel transformation suppressor, Pdcd4, inhibits AP-1 transactivation but not NF-kappaB or ODC transactivation. Oncogene. 20:669–676. 2001. View Article : Google Scholar : PubMed/NCBI
31	Vikhreva PN, Shepelev MV and Korobko IV: mTOR-dependent transcriptional repression of Pdcd4 tumor suppressor in lung cancer cells. Biochim Biophys Acta. 1839:43–49. 2014. View Article : Google Scholar : PubMed/NCBI