1.oxLDL FUNCTIONALLY UPREGULATES GROWTH-REGULATED ONCOGENE ? EXPRESSION IN HUMAN UMBILICAL VEIN ENDOTHELIAL CELLS
Zhubin LEI ; Jianping LIU ; Xiaoyan LI
Medical Journal of Chinese People's Liberation Army 2001;0(12):-
To study the effect of chemokine growth-regulated oncogene ? (GRO?) in process of LDL-induced atherosclerosis, a monocytic cell line, U937 cells and human umbilical vein endothelial cells were used. GRO? mRNA levels were significantly elevated after exposure of endothelial cells to oxLDL. A significant upregulation of GRO? surface expression in endothelial cells was induced by oxLDL with concentration-dependent and time-dependent manners, but no significant change in GRO? was observed in the medium. Upregulation of GRO? expression in endothelial cells enhanced monocytic cells adhesion to endothelial cells, but antibodies to GRO? inhibited the adhesion process. oxLDL functionally upregulates GRO? expression in endothelial cells.
2.Mechanism of the proliferation and apoptosis of keloid fibroblasts induced by 5-fluouracil:a preliminary study
Zhubin PAN ; Xiaojing LI ; Zongbao ZUO ; Chaohua LIU ; Chun WANG
Chinese Journal of Medical Aesthetics and Cosmetology 2009;15(5):323-326
Objective To investigate the of 5-fluorouracil effects on the expression of Smad7,TGF-β receptorⅠ,Bcl-2 and Bax in keloid fibroblasts.Methods After primary culture of keloid fibroblasts,4-6 passages of cells were inoculated in 5 different concentrations of 5-fluorouracil(10,20,40,80,160μmol/L)for 24,48 and 72 hours.Proliferative ability of keloid fibroblasts was detected by MTT assay.Expression of Smad 7,TGF-βreceptorⅠ,Bcl-2 and Bax in keloid fibroblasts was measured by Western blot.Results During MTT,5-fluorouracil did not affect cell viability at 24 hour at the concentration of 10 and 20 μmol/L.Compared with the control group,no significant difference was detected(P>0.05).At other concentrations,fibroblast death was visible in each group(P<0.01).Western blot analysis showed that the expression of Smad7 significantly decreased and the expression of TGF-β receptor Ⅰ significantly increased in the TGF-β1 group compared with the blank control group(P<0.0 1).5-fluorouracil could significantly enhance the expression of Smad7(P<0.01).There was a remarkable decrease of the Bcl-2 expression and marked increase of the Bax expression in different concentrations of 5-fluorouracil compared with the control group(P<0.05).But,5-fluorouracil did not show any effect on the synthesis of TGF-β receptor Ⅰ.Conclusion 5-fluorouracil could inhibit proliferation and induce apoptosis on human keloid fibroblasts in vitro.
3.Enrichment and isolation of breast cancer stem cells from breast cancer transplantation tumors in nude mice
Xintao LI ; Zhen HU ; Zhubin LI ; Qianqian GENG
Cancer Research and Clinic 2022;34(10):721-725
Objective:Enriching and isolating breast cancer stem cells from breast cancer transplantation tumors in nude mice.Methods:Human breast cancer MDA-MB-231 cells were injected into the right axilla subcutaneous of 20 nude mice, and the tumor growth was observed .After 30 days, tumors were isolated and stained with hematoxylin and eosin, and then tumor cells from tissues were isolated. DMEM medium containing serum was used to cultivate isolated transplantation tumor cells, cell morphology and growth were also observed. Flow cytometry was used to detect the proportion of stem cells (CD44 +/CD24 -/low cells) in transplantation tumor cells. Serum-free DMEM medium containing epidermal growth factor (EGF), basic fibroblast growth factor (bFGF) and B27 cell supplement were used to cultivate transplantation tumor cells and to obtain cell microspheres. The proportion of stem cells on the 10th day in cell microspheres was detected by using flow cell sorter and stem cells were isolated according to the markers of cell surface. Results:After subcutaneously injecting MDA-MB-231 cells into 20 nude mice for 9 days, 17 nude mice had subcutaneous tumors with more parenchymal cells, little interstitial cells, arranged cords tumor cells, large volume of the cell and abundant cytoplasm, the nuclei in different sizes and hyperchromatic state, mitotic more common, the nucleoli clear and obvious pleomorphy. After cultivating transplantation tumor cells with DMEM medium containing serum, the cells began to grow adherent after 24 h, and the adherent proportion rose to 60% after 3 days; after 7 days, the cell proliferation was accelerated; and the cell morphology was more consistent, most of which were spindle shaped and were not significantly different from MDA-MB-231 cells; the proportion of stem cells in transplantation tumor cells was (0.10±0.02)%. After cultivating transplantation tumor cells with serum-free DMEM medium containing cell cultured supplement, the cells grow in spherical patterns, the proportion of stem cells in cell microspheres got up to (70.47±2.03)% on the 10th day.Conclusions:Subcutaneously injecting MDA-MB-231 cells in nude mice can build breast cancer nude mice ectopic transplantation tumor model. Breast cancer stem cells in the transplantation tumors can be enriched from isolated transplantation tumor cells through serum-free medium, and more stem cells can be isolated to provide the research basis for the biological characteristics of breast cancer stem cells.