ABSTRACT
Objective To investigate the expression of Nestin and its relationship with tamoxifen cura-tive effect and prognosis of patients with breast cancer. Methods A total of 82 patients with breast cancer who received tamoxifen therapy after radical mastectomy in Department of Breast Surgery of Enshi Tujia Miao Auton-omous Prefecture Central Hospital of Hubei Province from March 2009 to March 2013 were collected. And the paired cancer tissues and adjacent normal tissues preserved in liquid nitrogen were also collected. Fluorescent quantitative real-time PCR( RT-PCR)and immunohistochemistry were used to detect the content of Nestin mRNA and the expression of Nestin in breast cancer tissues. The expression of Nestin in breast cancer tissues and its relationship with clinicopathologic features were analyzed. Breast cancer cell line MCF-7 was selected, small interfering RNA(siRNA)was used to silence Nestin in MCF-7 cells,and the influence on tamoxifen sen-sitivity was observed. The relationship of Nestin and epithelial-mesenchymal transition( EMT)was detected using Western blotting. Results The results of RT-PCR indicated that the mRNA level of Nestin in breast cancer tissues was 3. 87 times as high as that in paracancerous tissues(6. 34 ± 1. 56 vs. 1. 64 ± 0. 52,t =26. 140,P < 0. 001). Immunohistochemical staining suggested that the positive rate of Nestin in breast cancer tissues was 75. 61％(62 / 82),which was significantly higher than that in paracancerous tissues[24. 39％(20 / 82)],with a significant difference(χ2 = 43. 024,P < 0. 001). The expression of Nestin was related to lymphatic vessel infiltration(χ2 = 7. 499,P = 0. 006)and lymph node metastasis(χ2 = 6. 770,P = 0. 034), and it was not related to the age of patients(χ2 = 3. 242,P = 0. 072),tumor size(χ2 = 2. 358,P = 0. 308), histological grade(χ2 = 0. 294,P = 0. 863),ductal infiltrating status(χ2 = 0. 180,P = 0. 671)and triple neg-ative breast cancer(χ2 = 0. 142,P = 0. 706). The analysis of Cox risk ratio model showed that Nestin expres-sion(HR = 1. 982,P < 0. 001;HR = 1. 537,P < 0. 001),lymphatic vessel invasion( HR = 2. 502,P <0. 001;HR = 1. 715,P < 0. 001)and lymph node metastasis(HR = 1. 818,P < 0. 001;HR = 1. 446,P <0. 001)were independent prognostic risk factors for progress-free survival and overall survival in breast cancer patients. After Nestin knockout in breast cancer cell line MCF-7,the IC50 value of MCF-7 to tamoxifen de-creased from(48. 05 ± 2. 22)μmol/ L to(35. 59 ± 2. 92)μmol/ L,with a significant difference(t = 6. 489, P = 0. 003). Silencing Nestin significantly up-regulated E-cadherin(7. 21 ± 1. 15 vs. 1. 02 ± 0. 01,t = 6. 654, P = 0. 024)and down-regulated the mesenchyme index Vimentin(0. 17 ± 0. 08 vs. 1. 01 ± 0. 02,t = 25. 015, P < 0. 001)in MCF-7 cells. Conclusion Nestin is over-expressed in breast cancer,which is associated with reduced efficacy of tamoxifen. Nestin may be a new potential prognostic biomarker for breast cancer.
ABSTRACT
AIM:To investigate whether ginsenoside Rg1 attenuates high-fat diet ( HFD)-induced non-alcoho-lic fatty liver disease ( NAFLD) by improving β-oxidation.METHODS: SD rats ( n=60) were randomly divided into control group ( CON ) , HFD group, low-dose, medium-dose and high-dose ginsenoside Rg1 groups ( LDG, MDG and HDG) and positive drug ( sodium ursodeoxycholate) treatment group ( PDT) .High-fat diet was given for 8 weeks to suc-cessfully establish an NAFLD model.The animals were treated with the appropriate medications for 4 weeks and 8 weeks af-ter modeling, and sacrificed to collect the liver tissues for observing the pathologic changes with HE staining and for detec-ting liver functions and lipid levels.The expression of hepatic acyl-CoA synthetase 1 (CoASH1), carnitine acyltransferase I (CATI) and acyl-CoA oxidase 1 (ACOX1) at mRNA and protein levels was determined by RT-PCR and Western blot-ting.RESULTS:After 4-week treatment, the fatty infiltration of the liver tissues in PDT group, LDG group and MDG group was not attenuated except HDG group.After 8 weeks of treatment, a small number of fat particles was observed in PDT group and LDG group, while no infiltration of lipid droplet was found in MDG group and HDG group.Compared with HFD group, the levels of AST, ALT, AKP, TC, TG and LDL-C were significantly decreased after 4-week treatment in PDT group, LDG group, MDG group and HDG group (P<0.05), these indexes were further reduced after 8-week treatment. After 4-week treatment, HDL-C was significantly increased in the 4 treatment groups and almost restored to the level of CON group after 8-week treatment.The levels of CoASH1, CACTI and ACOX1 in the liver tissue of the 4 treatment groups were significantly increased after 4-week treatment (P<0.05) and much improved after 8-week treatment, and those in MDG group and HDG group were better than those in PDT group (P<0.05).CONCLUSION:Ginsenoside Rg1 regulatesβ-oxidation-related enzymes to improve the fat metabolism, thus playing a therapeutic role in liver injury in the rats with NAFLD.