Experimental Studies For Expressions And Significances Of P57^KIP2, Estrogen Receptor, Progesterone Receptor In Endometrial Carcinoma

I The research in human endometrial adenocarcinoma and differentendometrium tissuesObjective: To discuss the effects of p57KIP2, estrogen receptor(ER) and progesteronereceptor (PR) on the genesis and progression of endometrial carcinoma.Methods：Choiced100cases of endometrial adenocarcinoma (EA), and choiced20casesof endometrial proliferative phase,20cases of endometrial secretory phase,20cases ofendometrial proliferative lesions,20cases of endometrial intraepithelial neoplasia (EIN),then observed the histology morphous of different endometrium tissues in HE stain. Theexpression of p57KIP2, ER and PR proteins in EA and different endometrium tissues weredetected by immunohistochemical EliVision methods, respectively.Results：The highest expression of p57KIP2protein was in endometrial secretory phaseand the lowest expression of p57KIP2protein was in proliferative lesions. The expression ofp57KIP2protein in EA was lower than EIN and secretory phase, but higher than theexpression in proliferative lesions and proliferative phase, but only the expression ofp57KIP2protein in secretory phase compared with the expression of p57KIP2protein in EA,proliferative lesions and proliferative phase were significant differences, respectively (P<0.01). The highest expression of ER protein was in proliferative lesions and the lowestexpression of ER protein was in EA. The expression of ER protein showed significantdifference between proliferative lesions and EA, and between proliferative phase and EA(P<0.01). The expression of PR protein in proliferative lesions and proliferative phase wasthe highest. The expression of PR protein in EA was the lowest. But there was nosignificant differences among the expression of PR protein in all lesions(P>0.05). In EA,there was a correlation between the respective expression of p57KIP2protein, ER protein and PR protein with histological grade and clinical stage (P<0.05). There was a correlationbetween the respective expression of p57KIP2protein and PR protein with muscularinfiltration depth, and ER protein and PR protein with age(P<0.05).There was acorrelation between the respective expression of p57KIP2protein, ER protein and PRprotein with lymph node metastasis (P<0.05).There was positive correlation betweenexpression of p57KIP2and PR, and between expression of ER and PR (P<0.01).Conclusion：The abnormal expression of p57KIP2, ER and PR protein might be importantbiological markers for malignant transformation in endometrium tissue. Progestogen maypromote the high expression of PR protein and p57KIP2protein. They may have a role innegative regulation of the cell cycle and inhibition of the development and progression inEC, synergistically. Estrogen may have a negative role in the expression of p57KIP2protein.They may have a role in positive regulation of the cell cycle and enhancement of thedevelopment and progression in EC. p57KIP2, ER and PR protein may help clinical toestimate the prognosis of EC.II The research in different human endometrium cellsObjective: To discuss the effects of p57KIP2, ER and PR on the occurrence anddevelopment of EC.Methods: Human EC cells (well differentiation endometrial carcinoma cell Ishikawa,moderately differentiation endometrial carcinoma cell JEC) were Cultured, and selectednormal human endometrial carcinoma cell as control group. Observed the morphology ofdifferent endometrial cells in the inverted microscope and HE; The growth of differentendometrial cells were detected by growth curve and MTT test; The expression of p57KIP2,ER and PR proteins in different endometrial cells, were detected by Western blot (WB);The cell cycle and apoptosis of different endometrial cells were detected by flow cytometry(FCM); The expression of p57KIP2gene in different endometrial cells were detected by Realtime fluorescent quantitative PCR (Real-time PCR).Results:①The cell growth curve:the JEC, Ishikawa, ESC cells gone to the logarithmicgrowth phase the third day, gone to the plateau phase the sixth days, JEC cells into recession phase the tenth days, Ishikawa and ESC cells gone to recession phase the ninethdays.②MTT: JEC had the highest OD value, ESC had the lowest OD value and the ODvalue of Ishikawa was the middle. There was significant difference in OD values betweenESC and JEC in the first to eight days, between Ishikawa and JEC in thefist,second,forth,fifth days, between ESC and Ishikawa in the forth,seventh,eighth days(P＜0.05). Three different endometrium cells all gone to the logarithmic growth phase in thesecond to fifth days.②WB: The highest expression of p57KIP2protein was in Ishikawa andthe lowest expression of p57KIP2protein was in ESC (P>0.05). The highest expression ofER and PR proteins were in ESC and the lowest expression of ER and PR proteins were inJEC,the expression of ER and PR proteins in ESC compared with Ishikawa and JEC weresignificant differences, respectively (P<0.05).③FCM: JEC had the highest proportion inG0/G1phase, Ishikawa had the lowest proportion in G0/G1phase; Ishikawa had thehighest proportion in S phase, JEC had the lowest proportion in S phase; The cellproportion in G0/G1phase and S phase of ESC compared with Ishikawa and JEC,Ishikawa compared with JEC, the differences were significant(P<0.05); JEC had thehighest proportion in G2/M phase, ESC had the lowest proportion in G2/M phase, but onlyESC compared with Ishikawa and JEC were significant differences, respectively (P<0.05).The highest apoptosis rate was in ESC and the lowest apoptosis rate was in JEC (P>0.05).④The Real-time PCR: The expression of p57KIP2mRNA in JEC was the highest, theexpression of p57KIP2mRNA in ESC was the lowest. The expression of p57KIP2mRNA inESC compared with Ishikawa and JEC, Ishikawa compared with JEC, were significantdifferences, respectively (P<0.05).Conclusion: The p57KIP2, ER and PR proteins may be involved in the occurrence anddevelopment of EC. High levels of estrogen and progesterone may have combined effect todown regulate the expression of p57KIP2mRNA and protein in normal endometrial cell; Orthere may exist p57KIP2gene mutation, transcribe and translate abnormal p57KIP2protein,make the function of negative regulation on cell cycle lost, lead to occurrence of EC.