HO-3867对胃癌细胞增殖、凋亡、迁移和侵袭的影响及机制
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摘要
目的观察姜黄素类似物HO-3867对胃癌细胞株MGC-3867增殖、凋亡、迁移和侵袭的影响,并探讨其作用机制。方法取胃癌细胞株MGC-803,将细胞分为HO-3867低剂量组、HO-3867中剂量组、HO-3867高剂量组,分别以2、4、6μmol/L的HO-3867处理细胞,空白对照组以等量血清培养基培养。采用细胞增殖实验检测细胞增殖能力;采用细胞克隆形成实验检测细胞克隆形成能力;通过Transwell小室建立细胞侵袭和迁移模型,检测细胞迁移侵袭能力;采用流式细胞仪检测细胞凋亡情况;采用Western blotting法检测抑癌基因抗第10号染色体同源缺失性磷酸酶-张力蛋白(PTEN)表达,并检测迁移相关蛋白基质金属蛋白酶2(MMP-2)、MMP-9及凋亡相关蛋白B淋巴细胞瘤-2(Bcl-2)、B淋巴细胞瘤-2相关蛋白(Bax)表达。结果 HO-3867低、中、高剂量组细胞活力值均低于空白对照组,细胞克隆形成、侵袭、迁移穿膜细胞数均少于空白对照组,总凋亡率均高于空白对照组(P均<0. 05)。HO-3867低、中、高剂量组MMP-2、MMP-9、Bcl-2蛋白表达均低于空白对照组(P均<0. 05),Bax、PTEN、p-PTEN蛋白表达均高于空白对照组(P均<0. 05)。结论 HO-3867可显著抑制胃癌细胞株MGC-3867的增殖、侵袭和迁移,并促进其凋亡,其机制可能与调节PTEN、Bcl-2、Bax、MMP-2以及MMP-9等蛋白表达有关。
Objective To observe the effects of curcumin analogue HO-3867 on the proliferation,apoptosis,migration and invasion of gastric cancer cell line MGC-3867 and to explore its mechanism. Methods Gastric cancer cells MGC-803 were divided into three groups: low-dose,medium-dose and high-dose HO-3867 groups,which were treated with 2,4,and 6 μ mol/L HO-3867,respectively,and the cells in the control group were cultured in the same amount of serum medium. The proliferation of MGC-803 cells was detected by cell proliferation assay. The cell invasion and migration models were established by Transwell chamber,and the migration and invasion abilities of MGC-803 cells were detected. The apoptosis of MGC-803 cells was detected by flow cytometry. The expression of tumor suppressor gene phosphatase and tensin homolog deleted on chromosome ten( PTEN) and the expression levels of migration-related proteins matrix metalloproteinase 9( MMP-9) and MMP-2,and apoptosis-related proteins B-cell lymphoma-2( Bcl-2) and BCL2-associated X( Bax)were detected by Western blotting. Results The cell viability of HO-3867 in the low-dose,medium-dose and high-dose HO-3867 groups was lower than that of the control group( P < 0. 05). The number of colonies and transmembrane cells of invasion and migration in the low-dose,medium-dose and high-dose HO-3867 groups were lower than those in the control group,and the total apoptosis rate was higher than that in the control group( all P < 0. 05). The expression levels of MMP-2,MMP-9 and Bcl-2 in the low-dose,medium-dose and high-dose HO-3867 groups were lower than those in the control group( all P < 0. 05). The expression of Bax,PTEN and p-PTEN in low-dose,medium-dose and high-dose HO-3867 groups was higher than that of the control group( P < 0. 05). Conclusion HO-3867 can significantly inhibit the proliferation,invasion and migration,and promote the apoptosis of gastric cancer cell line MGC-3867 by regulating the expression of PTEN,Bcl-2,Bax,MMP-2,and MMP-9.
Objective To observe the effects of curcumin analogue HO-3867 on the proliferation,apoptosis,migration and invasion of gastric cancer cell line MGC-3867 and to explore its mechanism. Methods Gastric cancer cells MGC-803 were divided into three groups: low-dose,medium-dose and high-dose HO-3867 groups,which were treated with 2,4,and 6 μ mol/L HO-3867,respectively,and the cells in the control group were cultured in the same amount of serum medium. The proliferation of MGC-803 cells was detected by cell proliferation assay. The cell invasion and migration models were established by Transwell chamber,and the migration and invasion abilities of MGC-803 cells were detected. The apoptosis of MGC-803 cells was detected by flow cytometry. The expression of tumor suppressor gene phosphatase and tensin homolog deleted on chromosome ten( PTEN) and the expression levels of migration-related proteins matrix metalloproteinase 9( MMP-9) and MMP-2,and apoptosis-related proteins B-cell lymphoma-2( Bcl-2) and BCL2-associated X( Bax)were detected by Western blotting. Results The cell viability of HO-3867 in the low-dose,medium-dose and high-dose HO-3867 groups was lower than that of the control group( P < 0. 05). The number of colonies and transmembrane cells of invasion and migration in the low-dose,medium-dose and high-dose HO-3867 groups were lower than those in the control group,and the total apoptosis rate was higher than that in the control group( all P < 0. 05). The expression levels of MMP-2,MMP-9 and Bcl-2 in the low-dose,medium-dose and high-dose HO-3867 groups were lower than those in the control group( all P < 0. 05). The expression of Bax,PTEN and p-PTEN in low-dose,medium-dose and high-dose HO-3867 groups was higher than that of the control group( P < 0. 05). Conclusion HO-3867 can significantly inhibit the proliferation,invasion and migration,and promote the apoptosis of gastric cancer cell line MGC-3867 by regulating the expression of PTEN,Bcl-2,Bax,MMP-2,and MMP-9.
引文
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[12]Pena-Blanco A,Garcia-Saez AJ.Bax,Bak and beyond-mitochondrial performance in apoptosis[J].FEBS J,2018,285(3):416-431.
[13]Steck PA,Pershouse MA,Jasser SA,et al.Identification of a candidate tumour suppressor gene,MMAC1,at chromosome 10q23.3that is mutated in multiple advanced cancers[J].Nat Genet,1997,15(4):356-362.
[14]Salmena L,Carracedo A,Pandolfi PP.Tenets of PTEN tumor suppression[J].Cell,2008,133(3):403-414.
[15]Takei Y,Saga Y,Mizukami H,et al.Overexpression of PTEN in ovarian cancer cells suppresses i.p.dissemination and extends survival in mice[J].Mol Cancer Ther,2008,7(3):704-711.
[16]de la Iglesia N,Konopka G,Puram SV,et al.Identification of a PTEN-regulated STAT3 brain tumor suppressor pathway[J].Genes Dev,2008,22(4):449-462.
[2]Bixel K,Saini U,Kumar BH,et al.Targeting STAT3 by HO3867induces apoptosis in ovarian clear cell carcinoma[J].Int J Cancer,2017,141(9):1856-1866.
[3]Tierney BJ,Mc Cann GA,Naidu S,et al.Aberrantly activated pSTAT3-Ser727 in human endometrial cancer is suppressed by HO-3867,a novel STAT3 inhibitor[J].Gynecol Oncol,2014,135(1):133-141.
[4]Hu Y,Zhao C,Zheng H,et al.A novel STAT3 inhibitor HO-3867 induces cell apoptosis by reactive oxygen species-dependent endoplasmic reticulum stress in human pancreatic cancer cells[J].Anticancer Drugs,2017,28(4):392-400.
[5]Selvendiran K,Ahmed S,Dayton A,et al.HO-3867,a curcumin analog,sensitizes cisplatin-resistant ovarian carcinoma,leading to therapeutic synergy through STAT3 inhibition[J].Cancer Biol T-her,2011,12(9):837-845.
[6]Sullivan ND,Mackie JT,Miller RI,et al.First case of psittacine proventricular dilatation syndrome(macaw wasting disease)in Australia[J].Aust Vet J,1997,75(9):674.
[7]Rath KS,Naidu SK,Lata P,et al.HO-3867,a safe STAT3 inhibitor,is selectively cytotoxic to ovarian cancer[J].Cancer Res,2014,74(8):2316-2327.
[8]Tierney BJ,Mc Cann GA,Cohn DE,et al.HO-3867,a STAT3inhibitor induces apoptosis by inactivation of STAT3 activity in BRCA1-mutated ovarian cancer cells[J].Cancer Biol Ther,2012,13(9):766-775.
[9]Moran A,Iniesta P,Garcia-Aranda C,et al.Clinical relevance of MMP-9,MMP-2,TIMP-1 and TIMP-2 in colorectal cancer[J].Oncol Rep,2005,13(1):115-120.
[10]Murphy DA,Courtneidge SA.The ins and outs of podosomes and invadopodia:characteristics,formation and function[J].Nat Rev Mol Cell Biol,2011,12(7):413-426.
[11]Mazewski C,Liang K,Gonzalez de Mejia E.Comparison of the effect of chemical composition of anthocyanin-rich plant extracts on colon cancer cell proliferation and their potential mechanism of action using in vitro,in silico,and biochemical assays[J].Food Chem,2018,242:378-388.
[12]Pena-Blanco A,Garcia-Saez AJ.Bax,Bak and beyond-mitochondrial performance in apoptosis[J].FEBS J,2018,285(3):416-431.
[13]Steck PA,Pershouse MA,Jasser SA,et al.Identification of a candidate tumour suppressor gene,MMAC1,at chromosome 10q23.3that is mutated in multiple advanced cancers[J].Nat Genet,1997,15(4):356-362.
[14]Salmena L,Carracedo A,Pandolfi PP.Tenets of PTEN tumor suppression[J].Cell,2008,133(3):403-414.
[15]Takei Y,Saga Y,Mizukami H,et al.Overexpression of PTEN in ovarian cancer cells suppresses i.p.dissemination and extends survival in mice[J].Mol Cancer Ther,2008,7(3):704-711.
[16]de la Iglesia N,Konopka G,Puram SV,et al.Identification of a PTEN-regulated STAT3 brain tumor suppressor pathway[J].Genes Dev,2008,22(4):449-462.