Effects of Total Glycosides of <i>Cistanche deserticola</i> on Proliferation, Apoptosis and Expression of Wnt/<i>β</i>-Catenin Signaling Pathway Related Protein of HepG2 Cells

FENG Duo; WANG Jing; JIANG Yongjun; ZHOU Shiqi; DUAN Hao; GUO Yu; ZHAO Jian; YAN Wenjie

doi:10.13386/j.issn1002-0306.2022100297

Volume 44 Issue 20

Oct. 2023

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Science and Technology of Food Industry > 2023 > 44(20): 389-397. > DOI: 10.13386/j.issn1002-0306.2022100297

FENG Duo, WANG Jing, JIANG Yongjun, et al. Effects of Total Glycosides of Cistanche deserticola on Proliferation, Apoptosis and Expression of Wnt/β-Catenin Signaling Pathway Related Protein of HepG2 Cells[J]. Science and Technology of Food Industry, 2023, 44(20): 389−397. (in Chinese with English abstract). doi: 10.13386/j.issn1002-0306.2022100297.

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Effects of Total Glycosides of Cistanche deserticola on Proliferation, Apoptosis and Expression of Wnt/β-Catenin Signaling Pathway Related Protein of HepG2 Cells

1.
Beijing Key Laboratory of Bioactive Substances and Functional Food, College of Biochemical Engineering, Beijing Union University, Beijing 100023, China
2.
Institute of Food and Nutrition Development, Ministry of Agriculture and Rural Affairs, Beijing 100081, China
3.
Inner Mongolia Sankou Biotechnology Co., Ltd., Ordos 017000, China

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Received Date: October 31, 2022
Available Online: August 06, 2023

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Abstract

Abstract

To explore the inhibitory effect of total glycosides of Cistanche deserticola (TG) on HepG2 cells and its mechanism. In this paper, different concentrations (0, 3.5, 10.5, 21, 31.5, 42 μg/mL) TG were treated 24 h on HepG2 liver cancer cells, and the viability of HepG2 cells was detected using CCK8 assay. Hoechst 33342/PI double staining method and Annexin V-FITC/PI were used to detect HepG2 cells apoptosis. The phenomenon of cell migration was detected by cell migration assay. Meanwhile, cell cycle progression changes were detected by flow cytometry. And the expression of α-fetoprotein (AFP), β-catenin, Dishevelled (Dsh), GSK-3β were detected by Western blot. The results showed that TG could reduce the proliferation of HepG2 cells in a concentration dependent manner, with only 31.04% cell viability when TG was 42 μg/mL. In addition, TG could damage the cell structure and induce cell apoptosis, and the apoptosis rate could be as high as 32.44% by AV/PI detection. Moreover, TG could also promote cell necrosis, and limit cell migration. There were significant differences between the treated group and the control group (P<0.05 or P<0.01). Meanwhile, high concentration of TG could arrest HepG2 cells in S and G₂/M phase. Finally, compared with the control group, the relative expression of β-catenin and Dsh decreased, while GSK-3β increased in the TG treated groups. In conclusion, TG could inhibit HepG2 cell proliferation by affecting cell cycle progression, promoting apoptosis, and limiting cell migration. The mechanism might be through Wnt/β-catenin signaling pathway, which activated GSK-3β to degrade β-catenin to achieve liver cancer inhibition.

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References

[1]	冯朵, 何悦, 蒋勇军, 等. 肉苁蓉抗衰老功能的研究进展[J]. 食品安全质量检测学报,2021,12(11):4429−4437. [FENG D, HE Y, JIANG Y J, et al. Research progress on anti-aging function of Cistanches[J]. Journal of Food Safety & Quality,2021,12(11):4429−4437. doi: 10.19812/j.cnki.jfsq11-5956/ts.2021.11.018 FENG D, HE Y, JIANG Y J, et al. Research progress on anti-aging function of Cistanches[J]. Journal of Food Safety & Quality, 2021, 12(11): 4429-4437. doi: 10.19812/j.cnki.jfsq11-5956/ts.2021.11.018
[2]	MA G, CHEN J, WEI T, et al. Inhibiting roles of FOXA2 in liver cancer cell migration and invasion by transcriptionally suppressing microRNA103a-3p and activating the GREM2/LATS2/YAP axis[J]. Cytotechnology,2021,73:523−537. doi: 10.1007/s10616-021-00475-2
[3]	李梅青, 王康, 周鑫. 绿豆活性肽对HepG2肝癌细胞增殖的抑制作用[J]. 中国食品学报,2018,18(10):52−57. [LI M Q, WANG K, ZHOU X. Inhibition of mung bean peptides on proliferation in HepG2 cell[J]. Journal of Chinese Institute of Food Science and Technology,2018,18(10):52−57. doi: 10.16429/j.1009-7848.2018.10.008 LI M Q, WANG K, ZHOU X. Inhibition of mung bean peptides on proliferation in HepG2 cell[J]. Journal of Chinese Institute of Food Science and Technology, 2018, 18(10): 52-57. doi: 10.16429/j.1009-7848.2018.10.008
[4]	HU Y, WANG S, WU X, et al. Chinese herbal medicine-derived compounds for cancer therapy: A focus on hepatocellular carcinoma[J]. J Ethnopharmacol,2013,149(3):601−612. doi: 10.1016/j.jep.2013.07.030
[5]	YE Y, SONG Y, ZHUANG J, et al. Anticancer effects of echinacoside in hepatocellular carcinoma mouse model and HepG2 cells[J]. J Cell Physiol,2019,234(2):1880−1888. doi: 10.1002/jcp.27063
[6]	齐鑫鑫, 由淑萍, 何转霞, 等. 肉苁蓉苯乙醇苷对HepG2肝癌细胞增殖和凋亡的影响[J]. 新疆医科大学学报,2021,44(9):1041−1047. [QI X X, YOU S P, HE Z X, et al. Effect of Cistanche deserticola phenylethanol glycoside on proliferation and apoptosis of HepG2 cells in vitro[J]. Journal of Xinjiang Medical University,2021,44(9):1041−1047. QI X X, YOU S P, HE Z X, et al. Effect of Cistanche deserticola phenylethanol glycoside on proliferation and apoptosis of HepG2 cells in vitro[J]. Journal of Xinjiang Medical University, 2021, 44(9): 1041-1047.
[7]	谢玉婷. 松果菊苷诱导人肾癌786-O细胞凋亡及其机制的研究[D]. 包头: 内蒙古科技大学包头师范学院, 2020 XIE Y T. Effects of echniacoside on 786-O cells apoptosis and inductive mechanisms in vitro[D]. Baotou: Baotou Teachers' College, 2020.
[8]	韩一梅, 靳伟民, 曾会, 等. 松果菊苷对结肠癌SW480细胞体内外增殖、侵袭和转移的影响[J]. 广州中医药大学学报,2020,37(8):1542−1549. [HAN Y M, JIN W M, ZENG H, et al. Effects of echniacoside on proliferation, invasion and metastasis of colon cancer SW480 cells in vitro and in vivo[J]. Journal of Guangzhou University of Traditional Chinese Medicine,2020,37(8):1542−1549. HAN Y M, JIN W M, ZENG H, et al, Effects of echniacoside on proliferation, invasion and metastasis of colon cancer SW480 cells in vitro and in vivo[J]. Journal of Guangzhou University of Traditional Chinese Medicine, 2020, 37(8): 1542-1549.
[9]	胡琼, 由淑萍, 刘涛, 等. 肉苁蓉苯乙醇总苷抗肝癌作用的实验研究[J]. 癌变·畸变·突变,2018,30(3):194−199. [HU Q, YOU S P, LIU T, et al. An investigation on the anti-liver cancer effect of cistanche[J]. Carcinogenesis, Teratogenesis & Mutagenesis,2018,30(3):194−199. HU Q, YOU S P, LIU T, et al. An investigation on the anti-liver cancer effect of cistanche[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2018, 30(3): 194-199.
[10]	XU Q, QIN W, WU F Z, et al. Effect of Roucongrong (Herba Cistanches Deserticolae) decoction on the substantia nigra through Wnt/β-catenin signaling pathway in rats with Parkinson's disease induced by 6-hydroxydopamine hydrochloride[J]. Journal of Traditional Chinese Medicine,2021,41(5):762−770.
[11]	尹帅领, 王海波, 杨硕. 肉苁蓉多糖通过激活Wnt/β-catenin信号通路对6-HODA致帕金森病大鼠的神经保护作用[J]. 中西医结合心脑血管病杂志,2020,18(8):1227−1230. [YIN S L, WANG H B, YANG S. Neuroprotective effects of Cistanche deserticola polysaccharide on Parkinson's rats induced by 6-HODA caused by activating the Wntt/β-catenin signaling pathway[J]. Chinese Journal of Integrative Medicine on Cardio-Cerebrovascular Disease,2020,18(8):1227−1230. YIN S L, WANG H B, YANG S. Neuroprotective effects of Cistanche deserticola polysaccharide on Parkinson’s rats induced by 6-HODA caused by activating the Wntt/β-catenin signaling pathway[J]. Chinese Journal of Integrative Medicine on Cardio-Cerebrovascular Disease, 2020, 18(8): 1227-1230.
[12]	汤呈怀. 松果菊苷通过抑制Wnt/β-catenin信号通路发挥抗乳腺癌效应的作用机制研究[D]. 重庆: 重庆医科大学, 2020 TANG C H. Echinacoside inhibits breast cancer cells by suppressing Wnt/β-catenin signaling pathway[D]. Chongqing: Chongqing Medical University, 2020.
[13]	冯磊, 马秋玲, 石琳, 等. 松果菊苷通过抑制SOX4/Wnt/β-Catenin信号抑制急性髓系白血病细胞增殖[J]. 免疫学杂志,2020,36(2):138−142. [FENG L, MA Q L, SHI L, et al. Echinacein inhibits the proliferation of acute myeloid leukemia cells by suppressing SOX4/Wnt/beta-Catenin signal[J]. Immunological Journal,2020,36(2):138−142. FENG L, MA Q L, SHI L, et al. Echinacein inhibits the proliferation of acute myeloid leukemia cells by suppressing SOX4/Wnt/beta-Catenin signal[J]. Immunological Journal, 2020, 36(2): 138-142.
[14]	CLEVERS H, NUSSE R. Wnt/β-catenin signaling and disease[J]. Cell,2012,149(6):1192−1205. doi: 10.1016/j.cell.2012.05.012
[15]	RUSSELL J O, MONGA S P. Wnt/β-catenin signaling in liver development, homeostasis, and pathobiology[J]. Annu Rev Pathol,2018,13:351−378. doi: 10.1146/annurev-pathol-020117-044010
[16]	LIU C Y, CHEN K F, CHEN P J. Treatment of liver cancer[J]. Cold Spring Harb Perspect Med,2015,5(9):a021535. doi: 10.1101/cshperspect.a021535
[17]	WANG F, LI R, TU P, et al. Total glycosides of Cistanche deserticola promote neurological function recovery by inducing neurovascular regeneration via Nrf-2/Keap-1 pathway in MCAO/R rats[J]. Frontiers in Pharmacology 2020, 11, 236.
[18]	宋秋佳. 预知子提取物对人HepG2肝癌细胞黏附、迁移和侵袭的抑制作用及相关机制[D]. 上海: 上海中医药大学, 2019 SONG Q J. Inhibitory effect of Akebia Fruit extracts on adhesion, migration and invasison of human HepG2 hepatoma cells and related mechanisms[D]. Shanghai: Shanghai University of Traditional Chinese Medicine, 2019.
[19]	SUN Q, WANG J, LI Y, et al. Synthesis and evaluation of cytotoxic activities of artemisinin derivatives[J]. Chem Biol Drug Des,2017,90(5):1019−1028. doi: 10.1111/cbdd.13016
[20]	ZHAO Y M, SUN L N, ZHOU H Y, et al. Voltage-dependent potassium channels are involved in glutamate-induced apoptosis of rat hippocampal neurons[J]. Neurosci Lett,2006,398(1−2):22−27. doi: 10.1016/j.neulet.2005.12.073
[21]	GALLE P R, FOERSTER F, KUDO M, et al. Biology and significance of alpha-fetoprotein in hepatocelluar carcinoma[J]. Liver Int,2019,39(12):2214−2229. doi: 10.1111/liv.14223
[22]	KAMARAJAH S K, FRANKEL T L, SONNENDAY C, et al. Critical evaluation of the American Joint Commission on Cancer (AJCC) 8th edition staging system for patients with Hepatocellular Carcinoma (HCC): A surveillance, epidemiology, end results (SEER) analysis[J]. J Surg Oncol,2018,117(4):644−650. doi: 10.1002/jso.24908
[23]	EDGE S B, COMPTON C C. The American Joint Committee on cancer: The 7th edition of the AJCC cancer staging manual and the future of TNM[J]. Ann Surg Oncol,2010,17(6):1471−1474. doi: 10.1245/s10434-010-0985-4
[24]	VAUTHEY J N, LAUWERS G Y, ESNAOLA N F, et al. Simplified staging for hepatocellular carcinoma[J]. Journal of Clinical Oncology: Official Journal of the American Society of Clinical Oncology,2002,20(6):1527−1536. doi: 10.1200/JCO.2002.20.6.1527
[25]	ZHENG R, ZHANG S, ZENG H, et al. Cancer incidence and mortality in China, 2016[J]. Journal of the National Cancer Center,2022,2(1):1−9. doi: 10.1016/j.jncc.2022.02.002
[26]	ANWANWAN D, SINGH S K, SINGH S, et al. Challenges in liver cancer and possible treatment approaches[J]. Biochim Biophys Acta Rev Cancer,2020,1873(1):188314. doi: 10.1016/j.bbcan.2019.188314
[27]	CHAFFER C L, WEINBERG R A. A perspective on cancer cell metastasis[J]. Science,2011,331(6024):1559−1564. doi: 10.1126/science.1203543
[28]	ZHONG Z, YU J, VIRSHUP D M, et al. Wnts and the hallmarks of cancer[J]. Cancer Metastasis Rev,2020,39:625−645. doi: 10.1007/s10555-020-09887-6
[29]	SYAM S, ABDUL A B, SUKARI M A, et al. The growth suppressing effects of girinimbine on HepG2 involve induction of apoptosis and cell cycle arrest[J]. Molecules,2011,16(8):7155−7170. doi: 10.3390/molecules16087155
[30]	ZHANG X, LUO W, ZHAO W, et al. Isocryptotanshinone induced apoptosis and activated MAPK signaling in human breast cancer MCF-7 cells[J]. J Breast Cancer,2015,18(2):112−118. doi: 10.4048/jbc.2015.18.2.112
[31]	PONSELVI INDUJA M, EZHILARASAN D, ASHOK VARDHAN N. Evolvulus alsinoides methanolic extract triggers apoptosis in HepG2 cells[J]. Avicenna J Phytomed,2018,8(6):504−512.
[32]	YANG G, WANG S P, ZHONG L F, et al. 6-Gingerol induces apoptosis through lysosomal-mitochondrial axis in human hepatoma G2 cells[J]. Phytother Res,2012,26(11):1667−1673. doi: 10.1002/ptr.4632
[33]	李健, 郭庚, 范益民, 等. Dishevelled促进脑胶质瘤增殖的实验研究[J]. 中国药物与临床,2021,21(18):3077−3080. [LI J, GUO G, FAN Y M, et al. Dishevelled promotes proliferation of glioma: An experimental study[J]. Chinese Remedies & Clinics,2021,21(18):3077−3080. LI J, GUO G, FAN Y M, et al. Dishevelled promotes proliferation of glioma: An experimental study[J]. Chinese Remedies & Clinics, 2021, 21(18): 3077-3080.
[34]	FIEDLER M, MENDOZA-TOPAZ C, RUTHERFORD T J, et al. Dishevelled interacts with the DIX domain polymerization interface of Axin to interfere with its function in down-regulating β-catenin[J]. Proc Natl Acad Sci U S A,2011,108(5):1937−1942. doi: 10.1073/pnas.1017063108
[35]	石庆强, 左国伟, 冯子强, 等. 人参皂苷Rh_2通过激活GSK-3β降解β-catenin发挥抗肝癌作用研究[J]. 中草药,2016,47(18):3231−3238. [SHI Q Q, ZUO G W, FENG Z Q, et al. Study on anti-hepatoma effect of ginsenoside Rh2 and mechanism of degradation of β-catenin through activating GSK-3β[J]. Chinese Traditional and Herbal Drugs,2016,47(18):3231−3238. doi: 10.7501/j.issn.0253-2670.2016.18.016 SHI Q Q, ZUO G W, FENG Z Q, et al. Study on anti-hepatoma effect of ginsenoside Rh2 and mechanism of degradation of β-catenin through activating GSK-3β[J]. Chinese Traditional and Herbal Drugs, 2016, 47(18): 3231-3238. doi: 10.7501/j.issn.0253-2670.2016.18.016
[36]	HE S, TANG S. WNT/β-catenin signaling in the development of liver cancers[J]. Biomed Pharmacother,2020,132:110851. doi: 10.1016/j.biopha.2020.110851
[37]	HU P, CHENG B, HE Y, et al. Autophagy suppresses proliferation of HepG2 cells via inhibiting glypican-3/wnt/β-catenin signaling[J]. Onco Targets Ther,2018,11:193−200. doi: 10.2147/OTT.S150520
[38]	GAO Y X, YANG T W, YIN J M, et al. Progress and prospects of biomarkers in primary liver cancer (Review)[J]. Int J Oncol,2020,57(1):54−66.
[39]	TANGKIJVANICH P, ANUKULKARNKUSOL N, SUWANGOOL P, et al. Clinical characteristics and prognosis of hepatocellular carcinoma: Analysis based on serum alpha-fetoprotein levels[J]. J Clin Gastroenterol,2000,31(4):302−308. doi: 10.1097/00004836-200012000-00007
[40]	冯朵, 段昊, 吕燕妮, 等. 肉苁蓉在我国保健食品中的应用[J]. 食品科技,2021,46(12):76−81. [FENG D, DUAN H, LÜ Y N, et al. Application of Cistanche deserticola Ma in functional food in China[J]. Food Science and Technology,2021,46(12):76−81. doi: 10.3969/j.issn.1005-9989.2021.12.spkj202112013 FENG D, DUAN H, LÜ Y N, et al. Application of Cistanche deserticola Ma in functional food in China[J]. Food Science and Technology, 2021, 46(12): 76-81. doi: 10.3969/j.issn.1005-9989.2021.12.spkj202112013