Sann-Joong-Kuey-Jian-Tang Induces Autophagy in HepG2 Cells Via Regulation of the Phosphoinositide-3 Kinase/Akt/mammalian Target of Rapamycin and P38 Mitogen-activated Protein Kinase Pathways

Overview

Journal Mol Med Rep

Specialty Molecular Biology

Date 2015 Apr 8

PMID 25847489

Citations 6

Authors

Wan-Ling Chuang

Chin-Cheng Su

Ping-Yi Lin

Chi-Chen Lin

Yao-Li Chen

Affiliations

Soon will be listed here.

Abstract

Sann-Joong-Kuey-Jian-Tang (SJKJT), a traditional Chinese medicine, was previously reported to induce autophagy and inhibit the proliferation of the human HepG2 hepatocellular carcinoma cell line via an extrinsic pathway. In the present study, the effects of SJKJT-induced autophagy and the cytotoxic mechanisms mediating these effects were investigated in HepG2 cells. The cytotoxicity of SJKJT in the HepG2 cells was evaluated using a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. The results demonstrated that the half-maximal inhibitory concentration of SJKJT was 2.91 mg/ml at 24 h, 1.64 mg/ml at 48 h and 1.26 mg/ml at 72 h. The results of confocal fluorescence microscopy indicated that SJKJT resulted in the accumulation of green fluorescent protein-LC3 and vacuolation of the cytoplasm. Flow cytometric analysis revealed the accumulation of acidic vesicular organelles. Furthermore, western blot analysis, used to determine the expression levels of autophagy-associated proteins, demonstrated that the HepG2 cells treated with SJKJT exhibited LC3B-I/LC3B-II conversion, increased expression levels of Beclin, Atg-3 and Atg-5 and reduced expression levels of p62 and decreased signaling of the phosphoinositide-3 kinase/Akt/mammalian target of rapamycin and the p38 mitogen-activated protein kinase pathways. Taken together, these findings may assist in the development of novel chemotherapeutic agents for the treatment of malignant types of liver cancer.

Citing Articles

The 3,3'-dimethoxy-4,4'-dihydroxy-stilbene Triazole (STT) Inhibits Liver Cancer Cell Growth by Targeting Akt/mTOR Pathway.

Sun M, Bai J, Wang H, Zhou L, Li S Dokl Biochem Biophys. 2024; 517(1):277-284.

PMID: 39002015 DOI: 10.1134/S1607672923600537.

Coumarins and Gastrointestinal Cancer: A New Therapeutic Option?.

Banikazemi Z, Mirazimi S, Dashti F, Mazandaranian M, Akbari M, Morshedi K Front Oncol. 2021; 11:752784.

PMID: 34707995 PMC: 8542999. DOI: 10.3389/fonc.2021.752784.

C21 Fraction Refined from Induced Apoptosis is Enhanced by Suppression of Autophagy in Human Gastric Cell Lines.

Li K, Hao K, Zhang Y, Xu A, Wang Q, Du Y ACS Omega. 2020; 5(39):25156-25163.

PMID: 33043194 PMC: 7542599. DOI: 10.1021/acsomega.0c02748.

Hydroxypyridinone-Coumarin Inhibits the Proliferation of MHCC97 and HepG2 Human Hepatocellular Carcinoma Cells and Down-Regulates the Phosphoinositide-3 Kinase Pathway.

Cui X, Qin X Med Sci Monit. 2020; 26:e920785.

PMID: 32218414 PMC: 7133445. DOI: 10.12659/MSM.920785.

modulates hepatoma cells autophagy by regulating PI3K/AKT signaling pathway.

He J, Deng L, Liu H, Chen T, Chen S, Xia S Aging (Albany NY). 2019; 11(2):350-370.

PMID: 30696802 PMC: 6366968. DOI: 10.18632/aging.101737.

References

Plotnikov A, Zehorai E, Procaccia S, Seger R . The MAPK cascades: signaling components, nuclear roles and mechanisms of nuclear translocation. Biochim Biophys Acta. 2010; 1813(9):1619-33. DOI: 10.1016/j.bbamcr.2010.12.012. View

Li H, Jin X, Zhang Z, Xing Y, Kong X . Inhibition of autophagy enhances apoptosis induced by the PI3K/AKT/mTor inhibitor NVP-BEZ235 in renal cell carcinoma cells. Cell Biochem Funct. 2012; 31(5):427-33. DOI: 10.1002/cbf.2917. View

Cheng C, Lin Y, Su C . Sann-Joong-Kuey-Jian-Tang increases the protein expression of microtubule-associated protein II light chain 3 in human colon cancer colo 205 cells. Mol Med Rep. 2011; 2(5):707-11. DOI: 10.3892/mmr_00000160. View

Paglin S, Hollister T, Delohery T, Hackett N, McMahill M, Sphicas E . A novel response of cancer cells to radiation involves autophagy and formation of acidic vesicles. Cancer Res. 2001; 61(2):439-44. View

Shinojima N, Yokoyama T, Kondo Y, Kondo S . Roles of the Akt/mTOR/p70S6K and ERK1/2 signaling pathways in curcumin-induced autophagy. Autophagy. 2007; 3(6):635-7. DOI: 10.4161/auto.4916. View

Fan M, Chambers T . Role of mitogen-activated protein kinases in the response of tumor cells to chemotherapy. Drug Resist Updat. 2002; 4(4):253-67. DOI: 10.1054/drup.2001.0214. View

Aoki H, Takada Y, Kondo S, Sawaya R, Aggarwal B, Kondo Y . Evidence that curcumin suppresses the growth of malignant gliomas in vitro and in vivo through induction of autophagy: role of Akt and extracellular signal-regulated kinase signaling pathways. Mol Pharmacol. 2007; 72(1):29-39. DOI: 10.1124/mol.106.033167. View

Thorburn A . Apoptosis and autophagy: regulatory connections between two supposedly different processes. Apoptosis. 2007; 13(1):1-9. PMC: 2601595. DOI: 10.1007/s10495-007-0154-9. View

Miao Q, Bi L, Li X, Miao S, Zhang J, Zhang S . Anticancer effects of bufalin on human hepatocellular carcinoma HepG2 cells: roles of apoptosis and autophagy. Int J Mol Sci. 2013; 14(1):1370-82. PMC: 3565325. DOI: 10.3390/ijms14011370. View

10.

Chen Y, Yan M, Chien S, Kuo S, Chen D, Cheng C . Sann-Joong-Kuey-Jian-Tang inhibits hepatocellular carcinoma Hep-G2 cell proliferation by increasing TNF-α, Caspase-8, Caspase- 3 and Bax but by decreasing TCTP and Mcl-1 expression in vitro. Mol Med Rep. 2013; 7(5):1487-93. DOI: 10.3892/mmr.2013.1381. View

11.

Mei S, Gu H, Ward A, Yang X, Guo H, He K . p38 mitogen-activated protein kinase (MAPK) promotes cholesterol ester accumulation in macrophages through inhibition of macroautophagy. J Biol Chem. 2012; 287(15):11761-8. PMC: 3320924. DOI: 10.1074/jbc.M111.333575. View

12.

Cui Q, Tashiro S, Onodera S, Minami M, Ikejima T . Oridonin induced autophagy in human cervical carcinoma HeLa cells through Ras, JNK, and P38 regulation. J Pharmacol Sci. 2007; 105(4):317-25. DOI: 10.1254/jphs.fp0070336. View

13.

Meijer A, Codogno P . Regulation and role of autophagy in mammalian cells. Int J Biochem Cell Biol. 2004; 36(12):2445-62. DOI: 10.1016/j.biocel.2004.02.002. View

14.

Zhou J, Zhang W, Liang B, Casimiro M, Whitaker-Menezes D, Wang M . PPARgamma activation induces autophagy in breast cancer cells. Int J Biochem Cell Biol. 2009; 41(11):2334-42. PMC: 2753745. DOI: 10.1016/j.biocel.2009.06.007. View

15.

Cheng C, Lin Y, Su C . Sann-Joong-Kuey-Jian-Tang up-regulates the protein expression of Fas and TNF-α in colo 205 cells in vivo and in vitro. Mol Med Rep. 2011; 3(1):63-7. DOI: 10.3892/mmr_00000219. View

16.

Bincoletto C, Bechara A, Pereira G, Santos C, Antunes F, Peixoto da-Silva J . Interplay between apoptosis and autophagy, a challenging puzzle: new perspectives on antitumor chemotherapies. Chem Biol Interact. 2013; 206(2):279-88. DOI: 10.1016/j.cbi.2013.09.018. View

17.

Chang L, Karin M . Mammalian MAP kinase signalling cascades. Nature. 2001; 410(6824):37-40. DOI: 10.1038/35065000. View

18.

Kabeya Y, Mizushima N, Ueno T, Yamamoto A, Kirisako T, Noda T . LC3, a mammalian homologue of yeast Apg8p, is localized in autophagosome membranes after processing. EMBO J. 2000; 19(21):5720-8. PMC: 305793. DOI: 10.1093/emboj/19.21.5720. View

19.

Hu W, Chen S, Zhang J, Lou X, Zhou H . Dihydroartemisinin induces autophagy by suppressing NF-κB activation. Cancer Lett. 2013; 343(2):239-48. DOI: 10.1016/j.canlet.2013.09.035. View

20.

Wu Y, Tan H, Huang Q, Ong C, Shen H . Activation of the PI3K-Akt-mTOR signaling pathway promotes necrotic cell death via suppression of autophagy. Autophagy. 2009; 5(6):824-34. DOI: 10.4161/auto.9099. View