1.Effects and mechanisms of histone deacetylase inhibitors on invasion and metastasis of Her2 positive breast cancer cell
Jiang YIN ; Hao LIU ; Guopei ZHENG ; Yixue GU ; Zhimin HE
Journal of Chinese Physician 2017;19(2):177-181
Objective To investigate the effect of histone deacetylase inhibitor on Her2 positive breast cancer cell line BT474 and SKBR3 in apoptosis and metastasis.Methods Histone deacetylase inhibitor MS-275,suberoylanilide hydroxamic acid (SAHA)(4 μmol/L,and 50 μmol/iL,respectively) treated the cell lines BT474 and SKBR3 cells.Flow cytometer examined the apoptosis ratio.Transwell tested their metastatic activity.Western blot assay was performed to detect the associated proteins.Results SAHA and MS-275 inhibited the cell survival.The BT474 cell survival was (39 ± 11) %,(54 ± 8) %,and the SKBR3 survival was (62 ± 6) %,(71 ± 9) %,according to the fluorescence-activated cell sorting (FACS) result.SAHA and MS-275 induced the BT474 cell apoptosis 8.46± 0.28 (P <0.01),4.15 ± 0.71 (P <0.01) fold change,respectively;and upregulated the SKBR3 cell apoptosis ratio 5.51 ± 1.24 (P <0.01),4.04 ±0.69 (P <0.01) fold.The Transwell result showed that SAHA,MS-275 inhibited the Transwell ability of BT474 from the control 184.7 ± 18.8 to 104.3 ± 7.1,131.3 ±9.1 per view,and the SKBR3 from control 60 ± 16.7 per view to 14.3 ± 6.5,34.3 ± 8.7 per view.The Western blot result showed that SAHA,MS-275 inhibited the protein level of vimentin,Her2,β-catenin,histone deacetylase inhibitor (HDACi),and upregulated the acetylation level of histone 3.The E-caherin protein was regulated in BT474 and SKBR3 cells.Conclusions MS-275,SAHA can induce BT474 and SKBR3 apoptosis significantly,also inhibit their metastatic activity.
2.Relationship of hypoxia inducible factor 1? and vascular endothelial growth factor expression with angiogenesis in gastric carcinoma
Yixue GU ; Hanxian LI ; Yin KAN ; Hanqun WAN ; Zhihan TANG ; Tianshu GAO
Journal of Chinese Physician 2001;0(03):-
Objective To investigate the expression of hypoxia inducible factor 1?(HIF-1?),vascular endothelial growth factor(VEGF) and micro vessel density(MVD) in gastric carcinoma and to explore their correlation with clinical pathological features such as cancer invasion and metastasis.Methods Forty-eight samples of gastric carcinoma tissues were examined for the expression of HIF-1?,VEGF and CD34 by immunohistochemical method.Results The positive expression rates of HIF-1? and VEGF were 66.7% and 60.4% in gastric carcinoma respectively.The mean value of MVD was 42.5?14.7 in gastric carcinoma.The expressions of HIF-1?,VEGF and the value of MVD were significantly correlated with the depth of invasion,lymph node metastasis and TNM stage.The HIF-1? expression was positively correlated with VEGF expression and MVD value.Conclusion The overexpression of HIF-1?,VEGF and MVD consist in gastric carcinoma tissue.The HIF-1? expression is positively correlated with VEGF expression and MVD value.The overexpression of HIF-1?,VEGF and MVD value are closely related with invasion,metastasis and poor biological behavior of gastric carcinoma.
3.Comparison of effects of different fixation methods on the posterior pole of guinea pig eyeballs
Xiuyan ZHANG ; Yixue YIN ; Jike SONG ; Hongsheng BI
International Eye Science 2024;24(8):1207-1212
AIM: To avoid the problem of retinal dissection in guinea pig large eyeball tissue sections, different methods were used to optimize the fixation effect of the posterior pole of the eyeball.METHODS: A total of 75 normal guinea pigs(2 weeks old)were randomly divided into 5 large groups. Group A(1-3 small groups), the entire eyeball was fixed with FAS, Davidson fixative 1(D1), and Davidson fixative 2(D2)for 24 h; group B(4-6 small groups), the entire eyeball was fixed with FAS, D1, and D2 for 1 h, then cut the cornea and fix it in their respective fixatives for 2 h; group C(7-9 small groups), the eyeball was fixed in FAS, D1, and D2 for 1 h, divided into left and right halves along the direction of the optic nerve, and then placed them in their respective fixation solutions for 2 h; group D(10-12 small groups), after fixation for 3 h in FAS, D1, and D2, the eyeball was divided into left and right halves along the optic nerve direction; group E(13-15 small groups), the cornea was cut after fixation for 3 h in FAS, D1, and D2. Hematoxylin-eosin(HE)staining was used to compare the fixation effect on posterior eyeball in each group.RESULTS: After fixation, the surface of the eyeballs in groups, 1-6 and 11-15 was smooth and round, with a transparent and bright color. In groups 7-10, the eyeballs were sunken, wrinkled, and deformed. The HE staining showed that the eyeball morphology of groups 1, 5, 6, 14, and 15 was significantly better than the other groups, with a regular internal tissue structure. The eyeballs of the other groups were sunken and wrinkled, and the internal tissue was curled and tangled, with severe retinal detachment. In groups 1, 5, 6, 14, and 15, the retina, choroid, and sclera tissues of group 14 were closely connected, without obvious retinal detachment, rupture, or curling. The tissue structure was clear and visible, and the cells were arranged neatly.CONCLUSION: The fixation effect of cutting the cornea after fixing guinea pig eyeball with D1 fixative for 3 h is the most ideal, and this operation method is simple and suitable for studying the related structures of the posterior pole of the eye.