To investigate exogenous PTEN gene transfected human breast cancer cell line MDA-MD-468.Methods Using the lipofectamine 2000 transfection technique,wild type PTEN gene was transducted into an in vitro cultured highly ...To investigate exogenous PTEN gene transfected human breast cancer cell line MDA-MD-468.Methods Using the lipofectamine 2000 transfection technique,wild type PTEN gene was transducted into an in vitro cultured highly metastatic breast cancer cell line MDA-MD-468.After transfection,the cells were selected by G418.The resistant clones were chosen and expanded in DMEM culture medium.RT-PCR,immunohistochemical method and western blot were used to determine the expression of target genes.Results An anti-G418 cell clone was established and expanded in culture.The transfected PTEN gene MDA-MD-468 cells showed expression of PTEN mRNA and PTEN protein.Conclusion Human breast cancer cell line MDA-MB-468 established in this study expresses consistently exogenous PTEN genes.4 refs,6 figs.展开更多
目的:探讨miR-28-3p在三阴性乳腺癌(triple negative breast cancer,TNBC)组织和细胞系中的表达及其对MDA-MB-468细胞恶性生物学行为的影响。方法:收集2013年1月至2014年1月河北医科大学第四医院乳腺中心手术切除的、经病理证实的83例女...目的:探讨miR-28-3p在三阴性乳腺癌(triple negative breast cancer,TNBC)组织和细胞系中的表达及其对MDA-MB-468细胞恶性生物学行为的影响。方法:收集2013年1月至2014年1月河北医科大学第四医院乳腺中心手术切除的、经病理证实的83例女性TNBC患者的癌组织和癌旁组织标本,以及TNBC细胞系MDA-MB-468、HCC-1937、MDA-MB-231、MDA-MB-436、MDA-MB-453和人正常乳腺上皮细胞MCF10A,用qPCR检测组织和细胞系中miR-28-3p的表达水平并分析其表达与患者临床病理特征的相关性。用miR-28-3p抑制剂转染MDA-MB-468细胞后,用CCK-8、流式细胞术、细胞划痕和Transwell实验分别检测miR-28-3p抑制剂对MDA-MB-468细胞增殖、凋亡、侵袭和迁移能力的影响,用Western blotting检测MDA-MB-468细胞中桥接整合因子1(bridging integrator-1,BIN1)蛋白的表达水平。通过生物信息学工具预测miR-28-3p的靶基因BIN1,用双荧光素酶报告基因实验验证miR-28-3p对BIN1的调控作用。结果:TNBC组织及细胞系中miR-28-3p表达水平显著高于癌旁组织及MCF10A细胞(均P<0.01);83例TNBC组织中共有56例(67.47%)高表达miR-28-3p,其高表达与患者的Ki-67表达水平、肿瘤大小和TNM分期密切相关(均P<0.05或P<0.01)。与miR-NC组比较,miR-28-3p抑制剂组MDA-MB-468细胞增殖、侵袭和迁移能力降低,凋亡率升高(均P<0.05或P<0.01)。双荧光素酶报告基因和实验证实BIN1是miR-28-3p的靶基因,miR-28-3p抑制剂可上调MDA-MB-468细胞中BIN1蛋白的表达(P<0.05)。结论:miR-28-3p在TNBC组织及细胞中呈高表达状态,mi R-28-3p抑制剂上调BIN1表达进而抑制MDA-MB-468细胞的增殖、迁移和侵袭能力,并促进其凋亡。展开更多
AIM: To eluddate the relations between the myeloperoxidase -468G→A polymorphism and the development of duodenal ulcer (DU), and to investigate the impacts of this host genetic polymorphism on the histopathological fe...AIM: To eluddate the relations between the myeloperoxidase -468G→A polymorphism and the development of duodenal ulcer (DU), and to investigate the impacts of this host genetic polymorphism on the histopathological featuresof Helicobacter pylori ( H pylori)-related gastritis. METHODS: In a case-control study of 115 consecutive DU patients and 182 controls, the myeloperoxidase-468G→A polymorphism was genotyped. Additionally, gastric mucosal changes were examined according to the updated Sydney System.RESULTS: The two study groups differed in the distributionsof myelperoxidase genotypes (P= 0.008). All six individuals carrying myeloperoxidase A/A genotypes were in the DU group. The carriage of myeloperoxidase allele A and H pylori infection were associated with an increased risk of DU with odds ratios (OR) of 2.3 and 5.8, respectively. Thecombined risk of the carriage of myeloperoxidase allele A and H pylori infection for DU was 8.7 (95% CI, 3.5-21.8). In the H pylori-infected individuals, allele A carriers displayed higher bacterial density scores (P = 0.04) inthe antrum than did non-carriers.CONCLUSION: This work verifies for the first time the association of myeloperoxidase-468G→A polymorphism with antral H pyloridensity and DU disease. The mechanisms underlying this genetic polymorphism in developing DU disease merit further investigations.展开更多
3-(5'-hydroxymethyl-2'-furyl)-1-benzyl indazole(YC-1),the hypoxia-inducible factor-1 alpha(HIF-1α) inhibitor,suppresses tumor proliferation and metastasis by down-regulating HIF-1α expression under hypoxic c...3-(5'-hydroxymethyl-2'-furyl)-1-benzyl indazole(YC-1),the hypoxia-inducible factor-1 alpha(HIF-1α) inhibitor,suppresses tumor proliferation and metastasis by down-regulating HIF-1α expression under hypoxic conditions.Our previous studies demonstrated that YC-1 inhibited breast cancer cell proliferation under normoxic conditions.In the current study,we investigated the targets of YC-1 and mechanism of its action in MDA-MB-468 breast cancer cells.In the in vitro experiments,we found that YC-1 significantly inhibited MDA-MB-468 cell proliferation in normoxia and hypoxia.Under normoxic conditions,YC-1 induced apoptosis of MDA-MB-468 cells and blocked cell cycle in the G1 phase,and these effects were possibly related to caspase 8,p21,and p27 expression.RT-PCR and Western blotting results showed that YC-1 primarily inhibited HIF-1α at the mRNA and protein levels under hypoxic conditions,but suppressed the expression of epidermal growth factor receptor(EGFR) at the mRNA and protein levels under normoxic conditions.In vivo,YC-1 prolonged survival,increased survival rate,decreased tumor size and metastasis rate,and inhibited tissue EGFR and HIF-1α expression.However,YC-1 exerted no obvious effect on body weight.These results indicate that YC-1 inhibits the proliferation of MDA-MB-468 cells by acting on multiple targets with minimal side effects.Thus,YC-1 is a promising target drug for breast cancer.展开更多
文摘To investigate exogenous PTEN gene transfected human breast cancer cell line MDA-MD-468.Methods Using the lipofectamine 2000 transfection technique,wild type PTEN gene was transducted into an in vitro cultured highly metastatic breast cancer cell line MDA-MD-468.After transfection,the cells were selected by G418.The resistant clones were chosen and expanded in DMEM culture medium.RT-PCR,immunohistochemical method and western blot were used to determine the expression of target genes.Results An anti-G418 cell clone was established and expanded in culture.The transfected PTEN gene MDA-MD-468 cells showed expression of PTEN mRNA and PTEN protein.Conclusion Human breast cancer cell line MDA-MB-468 established in this study expresses consistently exogenous PTEN genes.4 refs,6 figs.
基金Supported by the grants from the Research Foundation of Kaohsiung Veterans General Hospital, Kaohsiung, Taiwan, China VGHKS9274 and the National Science Council, Taiwan, China NSC-92-2314B-075B-006
文摘AIM: To eluddate the relations between the myeloperoxidase -468G→A polymorphism and the development of duodenal ulcer (DU), and to investigate the impacts of this host genetic polymorphism on the histopathological featuresof Helicobacter pylori ( H pylori)-related gastritis. METHODS: In a case-control study of 115 consecutive DU patients and 182 controls, the myeloperoxidase-468G→A polymorphism was genotyped. Additionally, gastric mucosal changes were examined according to the updated Sydney System.RESULTS: The two study groups differed in the distributionsof myelperoxidase genotypes (P= 0.008). All six individuals carrying myeloperoxidase A/A genotypes were in the DU group. The carriage of myeloperoxidase allele A and H pylori infection were associated with an increased risk of DU with odds ratios (OR) of 2.3 and 5.8, respectively. Thecombined risk of the carriage of myeloperoxidase allele A and H pylori infection for DU was 8.7 (95% CI, 3.5-21.8). In the H pylori-infected individuals, allele A carriers displayed higher bacterial density scores (P = 0.04) inthe antrum than did non-carriers.CONCLUSION: This work verifies for the first time the association of myeloperoxidase-468G→A polymorphism with antral H pyloridensity and DU disease. The mechanisms underlying this genetic polymorphism in developing DU disease merit further investigations.
基金supported by a grant from Jilin Province Science and Technology Development Project(No.200905198)
文摘3-(5'-hydroxymethyl-2'-furyl)-1-benzyl indazole(YC-1),the hypoxia-inducible factor-1 alpha(HIF-1α) inhibitor,suppresses tumor proliferation and metastasis by down-regulating HIF-1α expression under hypoxic conditions.Our previous studies demonstrated that YC-1 inhibited breast cancer cell proliferation under normoxic conditions.In the current study,we investigated the targets of YC-1 and mechanism of its action in MDA-MB-468 breast cancer cells.In the in vitro experiments,we found that YC-1 significantly inhibited MDA-MB-468 cell proliferation in normoxia and hypoxia.Under normoxic conditions,YC-1 induced apoptosis of MDA-MB-468 cells and blocked cell cycle in the G1 phase,and these effects were possibly related to caspase 8,p21,and p27 expression.RT-PCR and Western blotting results showed that YC-1 primarily inhibited HIF-1α at the mRNA and protein levels under hypoxic conditions,but suppressed the expression of epidermal growth factor receptor(EGFR) at the mRNA and protein levels under normoxic conditions.In vivo,YC-1 prolonged survival,increased survival rate,decreased tumor size and metastasis rate,and inhibited tissue EGFR and HIF-1α expression.However,YC-1 exerted no obvious effect on body weight.These results indicate that YC-1 inhibits the proliferation of MDA-MB-468 cells by acting on multiple targets with minimal side effects.Thus,YC-1 is a promising target drug for breast cancer.
