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Lappaconitine sulfate induces apoptosis in human colon cancer HT-29 cells and down-regulates PI3K/AKT/GSK3β signaling pathway

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Abstract

Lappaconitine (LA), a diterpenoid alkaloid extracted from the roots of Aconitum sinomontanum Nakai, possesses strong central analgesic, local anesthetic, antifebric and anti-inflflammatory effect. Lappaconitine hydrobromide (LH) is used clinically to treat analgesia, but its clinical application is limited because of its poor solubility in water. Studies have suggested that lappaconitine sulfate (LS) is a promising pain reliever, and this particular form not only has readily water soluble, but also exhibits anti-cancer effects. However, the mechanism of LS anti-cancer activity is poorly understood. The aim of this study was intended to investigate the role of LS in apoptosis of human colon cancer HT-29 cells and explore the potential molecular mechanism. Cell proliferation was detected by CCK-8 assay and EdU proliferation assay. Cell morphological change was expressed by Hoechst 33258 staining assay. Expression of apoptosis related proteins were detected by western blot. The effect of LS on cell cycle was detected by flow cytometry. Experimental results showed that LS exhibited anti-proliferative activity and induced apoptosis in HT-29 cells in a dose-dependent manner. LS increased the expression of p53, Bax, cleaved-PARP, cleaved-caspase-3/7/9, and inhibited Bcl-2 expression. LS affected cyclin D1 and p21 expression and induced cell cycle arrest in G0/G1 phase. Additionally, LY294002 signifificantly abrogated the activation of p-PI3K, p-Akt and p-GSK3β. To summarize, these results demonstrated that LS was able to prevent cell proliferation probably via PI3K/Akt/GSK3β signaling pathway.

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Abbreviations

LA:

Lappaconitine

LS:

Lappaconitine sulfate

LH:

Lappaconitine hydrobromide

CCK-8:

Cell Counting Kit-8

PBS:

phosphatebuffered saline

FTIR:

fourier transform infrared spectroscopy

OD:

absorbance value

EdU:

5′-ethynyl-2′-deoxyuridine

SDS-PAGE:

sodium dodecyl sulfate polyacrylamide gel electrophoresis

PVDF:

polyvinylidene difluoride

References

  • Chen J, Duan Y, Zhang X, Ye Y, Ge B, Chen J (2015) Genistein induces apoptosis by the inactivation of the IGF-1R/p-Akt signaling pathway in MCF-7 human breast cancer cells. Food Funct 6:995–1000

    Article  CAS  PubMed  Google Scholar 

  • Cheng EH, Kirsch DG, Clem RJ, Ravi R, Kastan MB, Bedi A, Ueno K, Hardwick JM (1997) Conversion of BcI-2 to a Bax-like death effector by caspases. Science 278:1966–1968

    Article  CAS  PubMed  Google Scholar 

  • Dattaroy D, Seth RK, Sarkar S, Kimono D, Albadrani M, Chandrashekaran V, Hasson FA, Singh UP, Fan D, Nagarkatti M, Nagarkatti P, Diehl AM, Chatterjee S (2018) Sparstolonin B (SSnB) attenuates liver fibrosisvia a parallel conjugate pathway involving P53-P21 axis, TGF-beta signaling and focal adhesion that is TLR4 dependent. Eur J Pharmacol 841:33–48

    Article  CAS  PubMed  Google Scholar 

  • Decker P, Isenberg D, Muller S (2000) Inhibition of caspase-3-mediated Poly(ADP-ribose) Polymerase (PARP) apoptotic cleavage by human PARP autoantibodies and effect on cells undergoing apoptosis. J Biol Chem 275:9043–9046

    Article  CAS  PubMed  Google Scholar 

  • Ghanemi FZ, Belarbi M, Fluckiger A, Nani A, Dumont A, Rosny CD, Aboura I, Khan AS, Murtaza B, Benammar C (2017) Carob leaf polyphenols trigger intrinsic apoptotic pathway and induce cell cycle arrest in colon cancer cells. J Funct Foods 33:112–121

    Article  CAS  Google Scholar 

  • Gong EY, Shin YJ, Hwang IY, Kim JH, Kim SM, Moon JH, Shin JS, Lee DH, Hur DY, Jin DH, Hong SW, Lee WK, Lee WJ (2016) Combined treatment with vitamin C and sulindac synergistically induces p53- and ROS-dependent apoptosis in human colon cancer cells. Toxicol Lett 258:126–133

    Article  CAS  PubMed  Google Scholar 

  • Guo R, Overman M, Chatterjee D, Rashid A, Shroff S, Wang H, Katz MH, Fleming JB, Varadhachary GR, Abbruzzese JL, Wang H (2014) Aberrant expression of p53 p21, cyclin D1, and Bcl2 and their clinicopathological correlation in ampullary adenocarcinoma. Hum Pathol 45:1015–1023

    Article  CAS  PubMed  Google Scholar 

  • Han B, Jiang P, Li Z, Yu Y, Huang T, Ye X, Li X (2018) Coptisine-induced apoptosis in human colon cancer cells (HCT-116) is mediated by PI3K/Akt and mitochondrial-associated apoptotic pathway. Phytomedicine 48:152–160

    Article  CAS  PubMed  Google Scholar 

  • He M, ZhangY, Xie F, Dou X, Han M, Zhang H (2018) Role of PI3K/Akt/NF-κB and GSK-3β pathways in the rat model of cardiopulmonary bypass-related lung injury. Biomed Pharmacother 106:747–754

    Article  CAS  PubMed  Google Scholar 

  • Herman-Antosiewicz A, Singh SV (2004) Signal transduction pathways leading to cell cycle arrest and apoptosis induction in cancer cells by Allium vegetable-derived organosulfur compounds: a review. Mutat Res 555:121–131

    Article  CAS  PubMed  Google Scholar 

  • Huang X, Qiao Y, Zhou Y, Ruan Z, Kong Y, Li G, Xie X, Zhang J (2018) Ureaplasma, spp. lipid-associated membrane proteins induce human monocyte U937 cell cycle arrest through p53-independent p21 pathway. Int J Med Microbiol 308:819–828

    Article  CAS  PubMed  Google Scholar 

  • Kim AD, Han X, Piao MJ, Hewage SR, Hyun CL, Cho SJ, Hyun JW (2015) Esculetin induces death of human colon cancer cells via the reactive oxygen species-mediated mitochondrial apoptosis pathway. Environ Toxicol Pharmacol 39:982–989

    Article  CAS  PubMed  Google Scholar 

  • Kitagishi Y, Nakanishi A, Ogura Y, Matsuda S (2014) Dietary regulation of PI3K/AKT/GSK-3β pathway in Alzheimer’s disease. Alzheimers Res Ther 6:35–41

    Article  PubMed  PubMed Central  Google Scholar 

  • Kuo YC, Kuo PL, Hsu YL, Cho CY, Lin CC (2006) Ellipticine induces apoptosis through p53-dependent pathway in human hepatocellular carcinoma HepG2 cells. Life Sci 78:2550–2557

    Article  CAS  PubMed  Google Scholar 

  • Lin N, Xiao LY, Lin PY, Zhang D, Chen QW (2005) Experimental study on anti-tumor effects of lappaconitine hydrobromide. TCM Res 18:16–18

    Google Scholar 

  • Liu B, Chen, Cao G, Dong Z, Xu J, Luo T, Zhang S (2017) MicroRNA-27b inhibits cell proliferation in oral squamous cell carcinoma by targeting FZD7 and Wnt signaling pathway. Arch Oral Biol 83:92–96

    Article  CAS  PubMed  Google Scholar 

  • Livraghi L, Garber JE (2015) PARP inhibitors in the management of breast cancer: current data and future prospects. BMC Med 13:188–203

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Ma J, Chen X, Hou C, Zhu J, Han X, Zhang J, Guo H (2017) Effects of Lappaconitine sulphate on Hela cell proliferation, apoptosis and cell cycle. Chin. Pharm J 52:1038–1043

    CAS  Google Scholar 

  • Ma J, Han X, Chen X, Hou C, Zhu J, Yang C, Guo H (2017) Effects of Lappaconitine sulphate on HepG2 cell proliferation, apoptosis and cell cycle. Chinese Traditional Patent. Medicine 39:1940–1942

    Google Scholar 

  • Magiera K, Tomala M, Kubica K, De Cesare V, Trost M, Zieba BJ, Kachamakova-Trojanowska N, Les M, Dubin G, Holak TA, Skalniak L (2017) Lithocholic acid hydroxyamide destabilizes cyclin D1 and induces G0/G1 arrest by inhibiting deubiquitinase USP2a. Cell Chem Biol 24:458–470. e1–e18

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Martini M, De Santis MC, Braccini L, Gulluni F, Hirsch E (2014) PI3K/AKT signaling pathway and cancer: an updated review. Ann Med 46:372–383

    Article  CAS  PubMed  Google Scholar 

  • Peng L, Eltgroth ML, LaTempa TJ, Grimes CA, Desai TA (2009) The effect of TiO2 nanotubes on endothelial function and smooth muscle proliferation. Biomaterials 30:1268–1272

    Article  CAS  PubMed  Google Scholar 

  • Qian L, Liu Y, Xu Y, Ji W, Wu Q, Liu Y, Gao Q, Su C (2015) Matrine derivative WM130 inhibits hepatocellular carcinoma by suppressing EGFR/ERK/MMP-2 and PTEN/AKT signaling pathways. Cancer Lett 368:126–134

    Article  CAS  PubMed  Google Scholar 

  • Sandor V, Senderowicz A, Mertins S, Sackett D, Sausville E, Blagosklonny MV, Bates SE (2000) P21-dependent G1 arrest with downregulation of cyclin D1 and upregulation of cyclin E by the histone deacetylase inhibitor FR901228. Brit J Cancer 83:817–825

    Article  CAS  PubMed  Google Scholar 

  • Sheng LY, Xu M, Xu LQ, Xiong P (2013) Effect and mechanisms of lappaconitine on human lung cancer cells in vitro. J Chin Med Mater 5:840–843

    Google Scholar 

  • Sun C, Zhang Z, He P, Zhou Y, Xie X (2017) Involvement of PI3K/Akt pathway in the inhibition of hepatocarcinoma cell invasion and metastasis induced by SASH1 through downregulating Shh-Gli1 signaling. Int J Biochem Cell Biol 89:95–100

    Article  CAS  PubMed  Google Scholar 

  • Sun W, Zhang S, Wang H, Wang Y (2015) Synthesis, characterization and antinociceptive properties of the lappaconitine salts. Med Chem Res 24:3474–3482

    Article  CAS  Google Scholar 

  • Surachai M, Nisachon J, Jureerut D, Sompong K (2018) KT2 and RT2 modified antimicrobial peptides derived from Crocodylus siamensis Leucrocin I show activity against human colon cancer HCT-116 cells. Environ Toxicol Phar 62:164–176

    Article  CAS  Google Scholar 

  • Tang H, Wen FL, Wang SH, Liu XY, Chen DL, Wang FP (2016) New C20-diterpenoid alkaloids from Aconitum sinomontanum. Chin Chem Lett 27:761–763

    Article  CAS  Google Scholar 

  • Wang SQ, Hou HL, Bie LY, Nie CY, Wang LN, Gao SB, Hu TT, Chen XB (2019) Mechanistic studies of the apoptosis induced by the macrocyclic natural product tetrandrine in MGC 803 cells. Med Chem Res 28:107–115

    Article  CAS  Google Scholar 

  • Wang PD, Ma XM, Zhang HL, Yang YM, Yang YR, Wang HC, Lao AN (1997) Effect of lappaconitine on ECG in anesthetized rats and its anti arrhythmic. Action Chin Pharmacol Bull 13:73–75

    CAS  Google Scholar 

  • Wang J, Yuan L, Xiao H, Xiao C, Wang Y, Liu X (2013) Momordin Ic induces HepG2 cell apoptosis through MAPK and PI3K/Akt-mediated mitochondrial pathways. Apoptosis 18:751–765

    Article  CAS  PubMed  Google Scholar 

  • Wang YZ, Xiao YQ, Zhang C, Sun XM (2009) Study of analgesic and anti-inflammatory effects of lappaconitine gelata. J Trad Chin Med 29:141–145

    Article  Google Scholar 

  • Wei BY, Kong XW, Zhao ZY, Wang HC, Zhu RH (1981) Studies of Chinese Aconitum SPP, XVIII-alkaloids from Aconitum Sinomon-Tanum (1). Bull Chin Mater Med 6:26–28

    CAS  Google Scholar 

  • Wu YH, Ning YZ, Xu JB, Tan YH, Wu YY, Yan DA, Lǚ GQ (2008) A study on shenfu injection and lappaconitine hydrobromide injection inducing HL-60 differentiation and apoptosis. J Guangzhou Univ Trad Chin Med 25:131–137

    Google Scholar 

  • Xu H, Zhong H, Liu M, Xu C, Gao Y (2011) Lappaconitine-loaded microspheres for parenteral sustained release: effects of formulation variables and in vitro characterization. Pharmazie 66:654–661

    CAS  PubMed  Google Scholar 

  • Yan X, Wang L, Xia Y, Qiu Y, Tian X, Lv Y, Tian F, Song G, Wang T (2017) Fluoride induces apoptosis in H9c2 cardiomyocytes via the mitochondrial pathway. Chemosphere 182:159–165

    Article  CAS  PubMed  Google Scholar 

  • Yang S, Liu M, Zhao Q, Zhao H, Xue W, Yang S (2015a) Antiproliferative and apoptosis inducing effect of essential oil extracted from Cyrtomium fortumei (J.) Smith leaves. Med Chem Res 24:1644–1652

    Article  CAS  Google Scholar 

  • Yang SP, Zhang HY, Beier RC, Sun FF, Cao XY, Shen JZ, Wang ZH, Zhang SX (2015b) Comparative metabolism of Lappaconitine in rat and human liver microsomes and in vivo of rat using ultra high-performance liquid chromatography-quadrupole/time-of-flight mass spectrometry. J Pharm Biomed Anal 110:1–11

    Article  CAS  PubMed  Google Scholar 

  • Yü TT (1979) Aconitum sinomontanum Nakai. In: Flora of China. Science Press, Beijing, pp. 168–170

  • Yunusov MS (2011) Antiarrhythmic agents based on diterpenoid alkaloids. Russ Chem Bull 60:633–638

    Article  CAS  Google Scholar 

  • Zeng C, Ke ZF, Luo WR, Yao YH, Hu XR, Jie W, Yin JB, Sun SJ (2013) Heparanase overexpression participates in tumor growth of cervical cancer in vitro and in vivo. Med Oncol 30:403–409

    Article  CAS  PubMed  Google Scholar 

  • Zhang H, Li F, Pan Z, Wu Z, Wang Y, Cui Y (2014) Activation of PI3K/Akt pathway limits JNK-mediated apoptosis during EV71 infection. Virus Res 192:74–84

    Article  CAS  PubMed  Google Scholar 

  • Zhao XX, Chang JJ, Wang QL, Lu R, Li LJ, Sun X, Xie WD, Li X (2016) 5,6-Dihydroxy-3,7,4’-trimethoxyflavonol induces G2/M cell cycle arrest and apoptosis in human hepatocellular carcinoma cells. J Asian Nat Prod Res 18:1079–1090

    Article  CAS  PubMed  Google Scholar 

  • Zong A, Cao H, Wang F (2012) Anticancer polysaccharides from natural resources: a review of recent research. Carbohydr Polym 90:1395–1410

    Article  CAS  PubMed  Google Scholar 

  • Zuo T, Xu W, Li H, Song H, Zhu M (2017) Geniposide and geniposidic acid, modified forms of genipin, attenuate genipin-induced mitochondrial apoptosis without altering the anti-inflammatory ability in KGN cell line. Med Chem Res 26:499–508

    Article  CAS  Google Scholar 

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Acknowledgements

This work was supported by National Natural Science Foundation of China (No. 81760770 for Junyi Ma).

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Authors and Affiliations

  1. College of Life Science, Northwest Normal University, 730070, Lanzhou, Gansu, China

    Danni Qu, Xuemei Zhang, Yaqiong Zhou & Junyi Ma

  2. School of Pharmacy, Lanzhou University, 730000, Lanzhou, Gansu, China

    Chunyan Sang

  3. Experimental Center of Medicine, General Hospital of Lanzhou Military Command, 730050, Lanzhou, Gansu, China

    Ling Hui

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  1. Danni Qu
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Correspondence to Junyi Ma.

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Qu, D., Zhang, X., Sang, C. et al. Lappaconitine sulfate induces apoptosis in human colon cancer HT-29 cells and down-regulates PI3K/AKT/GSK3β signaling pathway. Med Chem Res 28, 907–916 (2019). https://doi.org/10.1007/s00044-019-02346-0

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