ABSTRACT
AIM:To investigate the effect of cis-dichlorodiamine platinun ( cDDP) resistance on proliferation , apoptosis, migration and angiogenesis of esophageal cancer cell line KYSE 150.METHODS:Using the method of increa-sing concentration of cDDP in culture for 10 months, the human esophageal carcinoma cDDP-resistant cell line named KYSE150/cDDP was established successfully .The drug sensitivity was measured by MTT assay .The changes of the biolog-ical behaviors between the parental cell line and resistant cell line were determined by morphological observation assay , MTT assay, colony formation assay , DAPI staining, wound healing assay and tube formation experiment .RESULTS: No significant morphological difference between KYSE 150 cells and KYSE150/cDDP cells was observed .Compared with KYSE150 cells, the drug resistance index of KYSE150/cDDP cells was 6.35, and the viability of KYSE150/cDDP cells was decreased.The colony formation rate of KYSE150/cDDP cells was (15.00 ±3.05)%, while the colony formation rate of KYSE150 cells was (86.70 ±6.57)%.The apoptotic rate of KYSE150/cDDP cells was (0.63 ±0.09)%, and that of KYSE150 cells was (8.46 ±1.33)%.Compared with KYSE150 cells, KYSE150/cDDP cells showed a stronger healing ability of scratch, and the migration rate was higher than that of KYSE 150 cells.The results of tube formation experiment showed that the vessel number in KYSE150/cDDP group was 76.20 ±3.18, while the vessel number in KYSE150 group was 50.60 ±1.33.The protein expression of MMP-2 and VEGFR2 in KYSE150/cDDP cells was higher than that in KYSE150 cells.CONCLUSION: KYSE150/cDDP cells present drug-resistant phenotype and show a slow growth rate . The ability of apoptosis is decreased , and the abilities of cell migration and angiogenesis are increased .This may be an im-portant reason for the failure of clinical chemotherapy for esophageal cancer .
ABSTRACT
Objective To study the expression and significance of Th17 and Tc17 cells in the peripheral blood,skin and lung in a murine model of bleomycin (BLM)-induced systemic sclerosis (SSc).Methods Thirty female BALB/c mouse were randomly divided into 3 groups,including a control group ( A group),a injected with BLM 4 week without pulmonary fibrosis(PF) group( B group) and with obviously PF group(C group).Pathological changes of skin and lung were detected.The proportion of CD4+,CD8+,CD4+IL-17+(Th17),CD8+IL-17+(Tc17) cells in the peripheral blood,skin and lung of mouse was determined by flow cytometry.The mRNA expressions of RORγt,IL-17A in skin and lung of mouse were evaluated by real-time PCR.Enzyme linked immunosorbent assay(ELISA) was used to measure the levels of IL-17 in serum.Results Dermal hydroxyproline(HYP) contents and the score of PF were significantly increased in C group [ (3.07±1.26) μg/mg,4.0±1.41 ]and B group [ (2.43±0.61) μμg/mg,1.50±0.76]as compared with A group [ (1.45±0.40) μg/mg,0.60±0.70 ],and pulmonary HYP contents was obviously increased in C group than in A and B groups,all P＜0.05.Compared with the A group,the percentage of CD4+ and Th17 cells in the peripheral blood,skin and lung of B and C groups,Tc17 cells of C group was significantly increased,and CD8+ cells was significantly decreased(all P＜0.05).The ratio of Th17/CD4+CD8+ in the peripheral blood,skin and lung of B and C groups [ ( 1.41 ±0.36)％,( 1.79±0.77)％ ],[ (2.58±1.07)％,(5.23±2.34)％ ]and [ (3.50±1.20)％,(4.02±1.32) ％ ]was significantly increased compared with A group (0.71±0.25)％,(1.15±0.59)％,(0.99±0.46)％.The ratio of Tc17/CD4+CD8+ in the lung of C groups( 1.62±0.53) ％ and in the skin of B and C groups [ (1.70±0.70) ％,( 1.63±0.63 ) ％ ]was significantly increased compared with A group [ ( 1.00±0.47 ) ％,( 1.1 1 ±0.34 ) ％ ],all P＜0.05.Compared with the A group,the mRNA levels of IL-17A,RORγt in skin of B and C groups,and in lung of C group were higher and the levels of IL-17 in serum was significantly increased,all P＜0.05.Th17 cells and the levels of IL-17 in blood were positive correlation with dermal and pulmonary inflammation,fibrosis and H YP contents,all P＜0.01.The frequency of Th17 and Tc17 cells in skin and lung respectively had a positive correlation with dermal and pulmonary inflammation,the score of fibrosis,and HYP contents of skin and lung,all P＜0.01.Conclusion Th17 and Tc17 cells were significantly increased in the peripheral blood,skin and lung of a murine model of SSc,and Th17 cells is dominated.They correlated with the inflammation and fibrosis of skin and lung,and may participate in the pathogenesis of SSc through secrete IL-17.