ABSTRACT
Objective To study the proliferation of human skin fibroblast and synthesis of extracellular matrix which were regulated by LATS1-YAP pathway. Methods They were divided into three groups:control groups, LATS1 siRNA intervention group and YAP siRNA treatment group. Using LATS1 siRNA transferred human skin fibroblasts cell lines HS27 in LATS1 siRNA intervention group, and using YAP siRNA transferred HS27 in YAP siRNA treatment group. Expression of LATS1,YAP and collageⅠwere detected by western-blot 48 h later, and the activity of HS27 cells was determined by MTT. Results Compared with control group,expression of LATS1 protein decreased while expression of YAP protein and collagenⅠprotein increased 48 h after LATS1 siRNA transfection. Expression of LATS1 protein remains un-changed and expression of YAP protein and collagenⅠprotein decreased 48 h after YAP siRNA transfection. Conclusion LATS1-YAP pathway could regulate proliferation of human skin fibroblast and synthesis of extracellular matrix. It provides a potential therapeutic targets for skin wound repair and cicatrization.
ABSTRACT
Objective To explore the biological effects of estrogen (17β-E2) on the proliferation and migration of human skin fibroblast (HSFB).Methods HSFBs were isolated and cultured by enzyme digestion.The fourth generation of HSFBs were adopted; (1) the proliferating effect of diverse concentrations of 17β-E2 and 17β-E2+ ICI-182780 on HSFBs was determined with MTT method at 24,48,72,96 h; (2) the influence of 17β-E2 and ICI-182780 to HSFBs cycle distribution were determined with flow cytometry; (3) the migration effect of diverse concentrations of 17β-E2 and 17β-E2+ICI-182780 on HSFBs was determined at 24,48,and 72 hours after the creation of the scratch-wound in vitro.Results (1) The proliferating speed of HSFBs in 10-10mol/L 17β-E2 group (group A)was the highest of all at 48,72,96 h,which was higher than that in ICI-182780+10-10mol/L 17β-E2 group (group B) and control group (group C) (P＜0.01) ;(2) the HSFBs during the S phase in group A was more than that in groups B and C (P＜0.01),while the HSFBs during the G0/G1 phase was less than that in groups B and C (P＜0.01); (3) the migrating effect of HSFBs in 10-8mol/L 17β-E2 group (group D) was the highest of all at 48 h,which was higher than that in ICI-182780+10-10mol/L control group (group E)and control group (group F) (P＜0.01).Conclusions The concentration of 10-10mol/L estrogen has the strongest effect of promoting proliferation and that of 10-8mol/L has the strongest chemotaxis; ICI-182780 can abate the above effect effectively.
ABSTRACT
PURPOSE: To investigate the percentage of colonies with 16 or more cells distribution of human skin fibroblast according to in vitro aging, and to evaluate the relationship between percentage of colonies with 16 or more cells and in vivo donor age in human skin fibroblast culture. MATERIAL AND METHOD: C1, C2, C3a, and C3b human skin fibroblast samples from three breast cancer patients were used as subjects. The C1, C2, and C3a donor were 44, 54, and 55 years old, respectively. C3a and C3b cells were isolated from the same person. Single cell suspension of skin fibroblasts was prepared with primary explant technique. One hundred cells are plated into 100ml tissue culture flask and cultured for two weeks. The colony size was defined as colonies with 16 or more cells. The cultured cell was stained with crystal violet, and number of cells in each colony was determined with stereo microscope at x10 magnification. Passage number of C1, C2, C3a and C3b skin fibroblast were 12th, 17th, and 14th, respectively. RESULTS: Percentage of colonies with 16 or more cells of skin fibroblast samples decreased with increasing in vitro passage number. In contrast, cumulative population doublings of skin fibroblast sample increased with increasing in vitro passage number. Percentage of colonies with 16 or more cells also decreased with increasing population doublings in human skin fibroblast culture. There was strong correlation with percentage of colonies with 16 or more cells and population doublings in C3a skin fibroblast sample. At the same point of population doublings, the percentage of colonies with 16 or more cells of the young C1 donor was higher level than the old C3a donor. CONCLUSION: The population doublings increased with increasing in vitro passage number but percentage of colonies with 16 or more cells decreased. The results of this study imply that percentage of colonies with 16 or more cells is useful as a indicator of in vitro human skin fibroblast aging and may estimate the in vivo donor age.