Effect of Yanggan Jiedu Sanjie formula on human hepatocellular carcinoma Bel-7402 cells
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摘要
OBJECTIVE: To observe the effect of Yanggan Jiedu Sanjie(YGJDSJ) formula on human hepatocellular carcinoma Bel-7402 cells.METHODS: Bel-7402 cells were treated with YGJDSJ. Cell proliferation was detected by cell counting kit-8 assay. Cell apoptosis was identified by Hoechst 33258 staining and flow cytometric analysis. Cell cycle distribution was quantified by flow cytometric analysis. Caspase activities were measured by commercial kit. Cell senescence was detected by senescence-associated β-galactosidase(SA-β-gal)staining. Protein expression and phosphorylation were identified by Western blot. Protein expression was knocked-down by siRNA.RESULTS: YGJDSJ inhibited proliferation of Bel-7402 cells in a dose-and time-dependent manner.YGJDSJ induced apoptosis and activated caspase-3, 8, and 9 in Bel-7402 cells. YGJDSJ-induced apoptosis was completely abrogated by a pan caspase inhibitor, Z-VAD-FMK. YGJDSJ also induced cell senescence, up-regulated cyclin-dependent kinase inhibitor 1 a(CDKN1 a) and CDKN2 a expression and down-regulated retinoblastoma protein(RB) phosphorylation in Bel-7402 cells. Specific knockdown of CDKN1 a and CDKN2 a significantly reduced YGJDSJ-induce cell senescence in Bel-7402 cells.CONCLUSION: YGJDSJ inhibited cell proliferation,induced caspase-dependent apoptosis and CDKN1 a/CDKN2 a-RB signalling mediated cell senescence in Bel-7402 cells. Our findings suggest that YGJDSJ might be potential for hepatocellular carcinoma treatment.
OBJECTIVE: To observe the effect of Yanggan Jiedu Sanjie(YGJDSJ) formula on human hepatocellular carcinoma Bel-7402 cells.METHODS: Bel-7402 cells were treated with YGJDSJ. Cell proliferation was detected by cell counting kit-8 assay. Cell apoptosis was identified by Hoechst 33258 staining and flow cytometric analysis. Cell cycle distribution was quantified by flow cytometric analysis. Caspase activities were measured by commercial kit. Cell senescence was detected by senescence-associated β-galactosidase(SA-β-gal)staining. Protein expression and phosphorylation were identified by Western blot. Protein expression was knocked-down by siRNA.RESULTS: YGJDSJ inhibited proliferation of Bel-7402 cells in a dose-and time-dependent manner.YGJDSJ induced apoptosis and activated caspase-3, 8, and 9 in Bel-7402 cells. YGJDSJ-induced apoptosis was completely abrogated by a pan caspase inhibitor, Z-VAD-FMK. YGJDSJ also induced cell senescence, up-regulated cyclin-dependent kinase inhibitor 1 a(CDKN1 a) and CDKN2 a expression and down-regulated retinoblastoma protein(RB) phosphorylation in Bel-7402 cells. Specific knockdown of CDKN1 a and CDKN2 a significantly reduced YGJDSJ-induce cell senescence in Bel-7402 cells.CONCLUSION: YGJDSJ inhibited cell proliferation,induced caspase-dependent apoptosis and CDKN1 a/CDKN2 a-RB signalling mediated cell senescence in Bel-7402 cells. Our findings suggest that YGJDSJ might be potential for hepatocellular carcinoma treatment.
OBJECTIVE: To observe the effect of Yanggan Jiedu Sanjie(YGJDSJ) formula on human hepatocellular carcinoma Bel-7402 cells.METHODS: Bel-7402 cells were treated with YGJDSJ. Cell proliferation was detected by cell counting kit-8 assay. Cell apoptosis was identified by Hoechst 33258 staining and flow cytometric analysis. Cell cycle distribution was quantified by flow cytometric analysis. Caspase activities were measured by commercial kit. Cell senescence was detected by senescence-associated β-galactosidase(SA-β-gal)staining. Protein expression and phosphorylation were identified by Western blot. Protein expression was knocked-down by siRNA.RESULTS: YGJDSJ inhibited proliferation of Bel-7402 cells in a dose-and time-dependent manner.YGJDSJ induced apoptosis and activated caspase-3, 8, and 9 in Bel-7402 cells. YGJDSJ-induced apoptosis was completely abrogated by a pan caspase inhibitor, Z-VAD-FMK. YGJDSJ also induced cell senescence, up-regulated cyclin-dependent kinase inhibitor 1 a(CDKN1 a) and CDKN2 a expression and down-regulated retinoblastoma protein(RB) phosphorylation in Bel-7402 cells. Specific knockdown of CDKN1 a and CDKN2 a significantly reduced YGJDSJ-induce cell senescence in Bel-7402 cells.CONCLUSION: YGJDSJ inhibited cell proliferation,induced caspase-dependent apoptosis and CDKN1 a/CDKN2 a-RB signalling mediated cell senescence in Bel-7402 cells. Our findings suggest that YGJDSJ might be potential for hepatocellular carcinoma treatment.
引文
1 Balogh J,Victor D 3rd,Asham EH,et al.Hepatocellular carcinoma:a review.J Hepatocell Carcinoma 2016;3:41-53.
2 Gong XL,Qin SK.Progress in systemic therapy of advanced hepatocellular carcinoma.World J Gastroenterol2016;22(29):6582-6594.
3 Torre LA,Bray F,Siegel RL,et al.Global cancer statistics,2012.CA Cancer J Clin 2015;65(2):87-108.
4 Hu B,Wang SS,Du Q.Traditional Chinese Medicine for prevention and treatment of hepatocarcinoma:from bench to bedside.World J Hepatol 2015;7(9):1209-1232.
5 Du Q,Hu B,Shen KP.Pathogenesis and treatment of hepatocellular carcinoma in Traditional Chinese Medicine.Shi Jie Zhong Xi Yi Jie He Za Zi 2010;5(9):814-817.
6 Hu B,An HM,Wang SS,et al.Preventive and therapeutic effects of Chinese herbal compounds against hepatocellular carcinoma.Molecules 2016;21(2):142.
7 Hu B,An HM,Shen KP,et al.Liver Yin deficiency tonifying herbal extract induces apoptosis and cell senescence in Bel-7402 human hepatocarcinoma cells.Exp Ther Med2012;3(1):80-86.
8 Hu B,Du Q,Deng S,et al.Ligustrum lucidum Ait.fruit extract induces apoptosis and cell senescence in human hepatocellular carcinoma cells through upregulation of p21.Oncol Rep 2014;32(3):1037-1042.
9 Hu B,Shen KP,An HM,et al.Aqueous extract of Curcuma aromatica induces apoptosis and G2/M arrest in human colon carcinoma LS-174-T cells independent of p53.Cancer Biother Radiopharm 2011;26(1):97-104.
10 Hu B,An HM,Shen KP,et al.Modified Yi Guan Jian,a Chinese herbal formula,induces anoikis in Bel-7402 human hepatocarcinoma cells in vitro.Oncol Rep 2011;26(6):1465-1470.
11 Hu B,An HM,Shen KP,et al.Polygonum cuspidatum extract induces anoikis in hepatocarcinoma cells associated with generation of reactive oxygen species and downregulation of focal adhesion kinase.Evid Based Complement Alternat Med 2012;2012:607675.
12 Koff JL,Ramachandiran S,Bernal-Mizrachi L.A time to kill:targeting apoptosis in cancer.Int J Mol Sci 2015;16(2):2942-2955.
13 Chiantore MV,Vannucchi S,Mangino G,et al.Senescence and cell death pathways and their role in cancer therapeutic outcome.Curr Med Chem 2009;16(3):287-300.
14 Schmitt CA.Cellular senescence and cancer treatment.Biochim Biophys Acta 2007;1775(1):5-20.
15 Lin HM,Tseng HC,Wang CJ,et al.Induction of autophagy and apoptosis by the extract of Solanum nigrum Linn in HepG2 cells.J Agric Food Chem 2007;55(9):3620-3628.
16 Wang HC,Chung PJ,Wu CH,et al.Solanum nigrum L.polyphenolic extract inhibits hepatocarcinoma cell growth by inducing G2/M phase arrest and apoptosis.J Sci Food Agric 2011;91(1):178-185.
17 Peng B,Hu Q,Liu X,et al.Duchesnea phenolic fraction inhibits in vitro and in vivo growth of cervical cancer through induction of apoptosis and cell cycle arrest.Exp Biol Med(Maywood)2009;234(1):74-83.
18 Cheng J,Han W,Wang Z,et al.Hepatocellular carcinoma growth is inhibited by Euphorbia helioscopia L.extract in nude mice xenografts.Biomed Res Int 2015;2015:601015.
19 Wang ZY,Liu HP,Zhang YC,et al.Anticancer potential of Euphorbia helioscopia L.extracts against human cancer cells.Anat Rec(Hoboken)2012;295(2):223-233.
20 Wang X,Jiang Y,Hu D.Optimization and in vitro antiproliferation of Curcuma Wenyujin's active extracts by ultrasonication and response surface methodology.Chem Cent J 2016;10:32.
21 Jiang S,Ling C,Li W,et al.Molecular mechanisms of anti-cancer activities ofβ-elemene:targeting hallmarks of cancer.Anticancer Agents Med Chem 2016;16(11):1426-1434.
22 Kasi PD,Tamilselvam R,Skalicka-Wo?niak K,et al.Molecular targets of curcumin for cancer therapy:an updated review.Tumour Biol 2016;37(10):13017-13028.
23 Du Q,Hu B,An HM,et al.Synergistic anticancer effects of curcumin and resveratrol in Hepa1-6 hepatocellular carcinoma cells.Oncol Rep 2013;29(5):1851-1858.
24 Shin JA,Shim JH,Jeon JG,et al.Apoptotic effect of Polygonum Cuspidatum in oral cancer cells through the regulation of specificity protein 1.Oral Dis 2011;17(2):162-170.
25 Lee CC,Chen YT,Chiu CC,et al.Polygonum cuspidatum extracts as bioactive antioxidaion,anti-tyrosinase,immune stimulation and anticancer agents.J Biosci Bioeng2015;119(4):464-469.
26 Carter LG,D'Orazio JA,Pearson KJ.Resveratrol and cancer:focus on in vivo evidence.Endocr Relat Cancer 2014;21(3):R209-R225.
27 Veena MS,Wilken R,Zheng JY,et al.p16 Protein and gigaxonin are associated with the ubiquitination of NFκBin cisplatin-induced senescence of cancer cells.J Biol Chem 2014;289(50):34921-3437.
28 Yang MY,Lin PM,Liu YC,et al.Induction of cellular senescence by doxorubicin is associated with upregulated miR-375 and induction of autophagy in K562 cells.PLoSOne 2012;7(5):e37205.
29 Han Z,Wei W,Dunaway S,et al.Role of p21 in apoptosis and senescence of human colon cancer cells treated with camptothecin.J Biol Chem 2002;277(19):17154-17160.
30 Heiss EH,Schilder YD,Dirsch VM.Chronic treatment with resveratrol induces redox stress-and ataxia telangiectasia-mutated(ATM)-dependent senescence in p53-positive cancer cells.J Biol Chem 2007;282(37):26759-26766.
31 Chang UM,Li CH,Lin LI,et al.Ganoderiol F,a ganoderma triterpene,induces senescence in hepatoma HepG2cells.Life Sci 2006;79(12):1129-1139.
32 Yu JH,Cui Q,Jiang YY,et al.Pseudolaric acid B induces apoptosis,senescence,and mitotic arrest in human breast cancer MCF-7.Acta Pharmacol Sin 2007;28(12):1975-1983.
33 Liu L,Wang J,Shi L,et al.β-Asarone induces senescence in colorectal cancer cells by inducing lamin B1 expression.Phytomedicine 2013;20(6):512-520.
34 Peng CC,Chen KC,Peng RY,et al.Antrodia camphorata extract induces replicative senescence in superficial TCC,and inhibits the absolute migration capability in invasive bladder carcinoma cells.J Ethnopharmacol 2007;109(1):93-103.
2 Gong XL,Qin SK.Progress in systemic therapy of advanced hepatocellular carcinoma.World J Gastroenterol2016;22(29):6582-6594.
3 Torre LA,Bray F,Siegel RL,et al.Global cancer statistics,2012.CA Cancer J Clin 2015;65(2):87-108.
4 Hu B,Wang SS,Du Q.Traditional Chinese Medicine for prevention and treatment of hepatocarcinoma:from bench to bedside.World J Hepatol 2015;7(9):1209-1232.
5 Du Q,Hu B,Shen KP.Pathogenesis and treatment of hepatocellular carcinoma in Traditional Chinese Medicine.Shi Jie Zhong Xi Yi Jie He Za Zi 2010;5(9):814-817.
6 Hu B,An HM,Wang SS,et al.Preventive and therapeutic effects of Chinese herbal compounds against hepatocellular carcinoma.Molecules 2016;21(2):142.
7 Hu B,An HM,Shen KP,et al.Liver Yin deficiency tonifying herbal extract induces apoptosis and cell senescence in Bel-7402 human hepatocarcinoma cells.Exp Ther Med2012;3(1):80-86.
8 Hu B,Du Q,Deng S,et al.Ligustrum lucidum Ait.fruit extract induces apoptosis and cell senescence in human hepatocellular carcinoma cells through upregulation of p21.Oncol Rep 2014;32(3):1037-1042.
9 Hu B,Shen KP,An HM,et al.Aqueous extract of Curcuma aromatica induces apoptosis and G2/M arrest in human colon carcinoma LS-174-T cells independent of p53.Cancer Biother Radiopharm 2011;26(1):97-104.
10 Hu B,An HM,Shen KP,et al.Modified Yi Guan Jian,a Chinese herbal formula,induces anoikis in Bel-7402 human hepatocarcinoma cells in vitro.Oncol Rep 2011;26(6):1465-1470.
11 Hu B,An HM,Shen KP,et al.Polygonum cuspidatum extract induces anoikis in hepatocarcinoma cells associated with generation of reactive oxygen species and downregulation of focal adhesion kinase.Evid Based Complement Alternat Med 2012;2012:607675.
12 Koff JL,Ramachandiran S,Bernal-Mizrachi L.A time to kill:targeting apoptosis in cancer.Int J Mol Sci 2015;16(2):2942-2955.
13 Chiantore MV,Vannucchi S,Mangino G,et al.Senescence and cell death pathways and their role in cancer therapeutic outcome.Curr Med Chem 2009;16(3):287-300.
14 Schmitt CA.Cellular senescence and cancer treatment.Biochim Biophys Acta 2007;1775(1):5-20.
15 Lin HM,Tseng HC,Wang CJ,et al.Induction of autophagy and apoptosis by the extract of Solanum nigrum Linn in HepG2 cells.J Agric Food Chem 2007;55(9):3620-3628.
16 Wang HC,Chung PJ,Wu CH,et al.Solanum nigrum L.polyphenolic extract inhibits hepatocarcinoma cell growth by inducing G2/M phase arrest and apoptosis.J Sci Food Agric 2011;91(1):178-185.
17 Peng B,Hu Q,Liu X,et al.Duchesnea phenolic fraction inhibits in vitro and in vivo growth of cervical cancer through induction of apoptosis and cell cycle arrest.Exp Biol Med(Maywood)2009;234(1):74-83.
18 Cheng J,Han W,Wang Z,et al.Hepatocellular carcinoma growth is inhibited by Euphorbia helioscopia L.extract in nude mice xenografts.Biomed Res Int 2015;2015:601015.
19 Wang ZY,Liu HP,Zhang YC,et al.Anticancer potential of Euphorbia helioscopia L.extracts against human cancer cells.Anat Rec(Hoboken)2012;295(2):223-233.
20 Wang X,Jiang Y,Hu D.Optimization and in vitro antiproliferation of Curcuma Wenyujin's active extracts by ultrasonication and response surface methodology.Chem Cent J 2016;10:32.
21 Jiang S,Ling C,Li W,et al.Molecular mechanisms of anti-cancer activities ofβ-elemene:targeting hallmarks of cancer.Anticancer Agents Med Chem 2016;16(11):1426-1434.
22 Kasi PD,Tamilselvam R,Skalicka-Wo?niak K,et al.Molecular targets of curcumin for cancer therapy:an updated review.Tumour Biol 2016;37(10):13017-13028.
23 Du Q,Hu B,An HM,et al.Synergistic anticancer effects of curcumin and resveratrol in Hepa1-6 hepatocellular carcinoma cells.Oncol Rep 2013;29(5):1851-1858.
24 Shin JA,Shim JH,Jeon JG,et al.Apoptotic effect of Polygonum Cuspidatum in oral cancer cells through the regulation of specificity protein 1.Oral Dis 2011;17(2):162-170.
25 Lee CC,Chen YT,Chiu CC,et al.Polygonum cuspidatum extracts as bioactive antioxidaion,anti-tyrosinase,immune stimulation and anticancer agents.J Biosci Bioeng2015;119(4):464-469.
26 Carter LG,D'Orazio JA,Pearson KJ.Resveratrol and cancer:focus on in vivo evidence.Endocr Relat Cancer 2014;21(3):R209-R225.
27 Veena MS,Wilken R,Zheng JY,et al.p16 Protein and gigaxonin are associated with the ubiquitination of NFκBin cisplatin-induced senescence of cancer cells.J Biol Chem 2014;289(50):34921-3437.
28 Yang MY,Lin PM,Liu YC,et al.Induction of cellular senescence by doxorubicin is associated with upregulated miR-375 and induction of autophagy in K562 cells.PLoSOne 2012;7(5):e37205.
29 Han Z,Wei W,Dunaway S,et al.Role of p21 in apoptosis and senescence of human colon cancer cells treated with camptothecin.J Biol Chem 2002;277(19):17154-17160.
30 Heiss EH,Schilder YD,Dirsch VM.Chronic treatment with resveratrol induces redox stress-and ataxia telangiectasia-mutated(ATM)-dependent senescence in p53-positive cancer cells.J Biol Chem 2007;282(37):26759-26766.
31 Chang UM,Li CH,Lin LI,et al.Ganoderiol F,a ganoderma triterpene,induces senescence in hepatoma HepG2cells.Life Sci 2006;79(12):1129-1139.
32 Yu JH,Cui Q,Jiang YY,et al.Pseudolaric acid B induces apoptosis,senescence,and mitotic arrest in human breast cancer MCF-7.Acta Pharmacol Sin 2007;28(12):1975-1983.
33 Liu L,Wang J,Shi L,et al.β-Asarone induces senescence in colorectal cancer cells by inducing lamin B1 expression.Phytomedicine 2013;20(6):512-520.
34 Peng CC,Chen KC,Peng RY,et al.Antrodia camphorata extract induces replicative senescence in superficial TCC,and inhibits the absolute migration capability in invasive bladder carcinoma cells.J Ethnopharmacol 2007;109(1):93-103.