| Objective:To detect the expression levels of mRNA and protein in endometrial tissues of HIF-1αand PHD2 under normal and different pathological conditions,whether there is any difference between normal and different pathological conditions,and endometrial adenocarcinoma The expression level in tissues is associated with high risk factors associated with prognosis of endometrial cancer(eg histological differentiation,infiltration more than 1/2 myometrial thickness,invasion of vascular septum,tumor diameter greater than 2 cm,metastasis to lymph nodes and outside the uterus),explore whether there is a correlation.The aim is to increase the evaluation index for the diagnosis of endometrial cancer,to help the early diagnosis,disease assessment and postoperative follow-up monitoring of endometrial cancer,and to improve the prognosis of patients with endometrial cancer.Methods:In our operating room,we collected endometrial tissue from June 2017 to December 2018 in our department.Among them,30 cases of dysplasia endometrial tissue were derived from irregular curettage due to vaginal irregular bleeding.30 cases were normal.Endometrial tissue due to non-endometrial lesions for hysterectomy,cervical CINIII lesions(the endometrium is required to be in the secretory phase);50 cases of endometrial adenocarcinoma Patients who underwent staged endometrial adenocarcinoma(all cases are type I endometrial adenocarcinoma,endometrioid adenocarcinoma,all uterine double attachment resection,pelvic abdominal aortic para-aortic lymphadenectomy,treatment with Chinese medicine,hormones,radiotherapy and chemotherapy were considered as exclusion criteria).The normal endometrial tissue was set as the control group,and the endometrium(dysplasia and adenocarcinoma)tissues under different pathological conditions were set as the experimental group.qRT-PCR and Western Blot assays were used to detect the expression levels of mRNA and protein in endometrial tissues(fresh tissues)under normal and different pathological conditions of PHD2 and HIF-1α.The paraffin sections of the corresponding patients were taken.Immunohistochemistry(Envision two-step method)was used to detect the expression levels of PHD2 and HIF-1αprotein.Correlation analysis method Pearson was used to analyze whether the expression levels of HIF-1αand PHD2 mRNA were correlated.Image J was used to detect the gray value(protein content)of WB bands;Graphpad prism7.0 mapping software was used to process experimental data;statistical software SPSS20.0 analysis of experimental data;T test analysis of qRT-PCR data,X~2 test analysis of immunohistochemistry results,detection of normal and different pathological conditions in the endometrial tissue PHD2 and HIF-1αexpression levels between the difference and in endometrial adenocarcinoma tissue There was a correlation between the expression level and the risk factors affecting the prognosis of endometrial cancer(P<0.05 was considered statistically significant).Results:The results of qRT-PCR showed that the expression levels of HIF-1αin normal endometrium,endometrial atypical hyperplasia and endometrial adenocarcinoma were0.714±0.652,3.444±1.171,20.830±10.761,the expression of HIF-1αmRNA was increased in the atypical hyperplasia group compared with the normal endometrium group(P<0.05),while the increase in HIF-1αwas more significant in the endometrial cancer tissues(P<0.01).The expression levels of PHD2 in normal endometrium,endometrial atypical hyperplasia and endometrial adenocarcinoma were 5.936±1.935,2.674±1.625,0.645±0.542,the expression of PHD2 mRNA in atypical hyperplasia group compared with the normal endometrium group,the decrease was observed(P<0.05),while in the endometrial cancer tissues,the decrease of PHD2 was more obvious(P<0.01).Pearson correlation analysis showed a negative correlation between HIF-1αand PHD2 expression levels at mRNA level,r=-0.175,P<0.05(due to|r|<0.3,suggesting a weak negative correlation between HIF-1αand PHD2).Paired T-test results suggest that the detection of HIF-1αexpression in the extent of endometrial lesions is more predictive than PHD2(T=4.361,P<0.01).The target bands of HIF-1αand PHD2 were located at 120KDa and46KDa,respectively.The gray value(protein content)of the WB target band was detected by Image J.The results showed:normal endometrium,endometrial atypical hyperplasia,endometrium in the adenocarcinoma tissues,the relative gray values of HIF-1αprotein expression were 76.706±27.907,125.436±22.691,and 153.449±32.425.The difference between the two groups was statistically significant(P<0.05);the relative expression of PHD2 protein was relative.The gray values were 154.159±8.359,113.392±12.266,72.683±18.198,and the difference between the groups was statistically significant(P<0.05).The Envision two-step method showed that PHD2 was positively expressed in cytoplasm,and its positive expression rate was 86.7%,70.0%and 28.0%in normal endometrium,endometrial atypical hyperplasia and endometrial adenocarcinoma tissues.HIF-1αwas positively expressed in cytoplasm and nucleus,mainly cytoplasm,and counted.The positive expression rate was 16.7%、40.0%and 82.0%in normal endometrium,endometrial atypical hyperplasia and endometrial adenocarcinoma tissues,the difference was statistically significant(P<0.05).qRT-PCR,Western Blot and Immunocytochemistry(Envision two-step method)showed that the expression of HIF-1αmRNA and protein was increased in the endometrial atypical hyperplasia group compared with the normal endometrial group,but in the endometrial adenocarcinoma.The increase of HIF-1αin tissues was more obvious(P<0.05);the expression of PHD2 mRNA and protein decreased in the endometrial atypical hyperplasia group compared with the normal endometrial group,while the PHD2 decreased in endometrial adenocarcinoma tissues.The trend was even more pronounced(P<0.05).The expression levels of HIF-1αand PHD2 protein were correlated with endometrial cancer vascular invasion,postoperative FIGO stage and lymph node metastasis(P<0.05),but not correlated with tissue differentiation,depth of myometrial invasion and tumor diameter(P>0.05).Conclusion:HIF-1αand PHD2 may be involved in the pathological process of the endometrium;HIF-1αand PHD2 may promote the invasion and metastasis of endometrial cancer;There was a negative correlation between HIF-1αand PHD2 mRNA expression levels;HIF-1αhad predictive value for endometrial cancer changes compared with PHD2. |
PDF Full Text Request