Expression Of HIF-2α And VE-cadherin In Cervical Adenocarcinoma And Their Correlation Study Under Hypoxia Condition

bjective: To investigate expression of hypoxia-inducible factor 2 alpha(HIF-2α) and Vascular endothelia-cadherin(VE-cadherin) and the relationship between the two factors in cervical adenocarcinoma tissues with immunohistochemical technique.The two cobalt chloride(Co Cl2) being established the model of external hypoxia model to study expression of HIF-2α and VE-cadherin in human cervical adenocarcinoma Hela cells at m RNA and protein levels under hypoxia conditions induced by.To explore under hypoxia conditions the effect of HIF-2α and VE-cadherin for the growth, invasion and metastasis of cervical adenocarcinoma, and the correlation between them. Methods : 1. Collected 60 cases cervical adenocarcinoma tissues and 18 cases normal cervical tissues screened to meet the standard, and analyse the expression of HIF-2α and VE-cadherin in tumor tissues by immunohistochemical technique, and make a further analysis on their clinical significance and their relationship. 2. Co Cl2 reagent established the model of external hypoxia model. 3. Human cervical adenocarcinoma Hela cells were cultured in vitro. After interventing with different concentrations of Co Cl2(50, 100, 150, 200,250μmol/L)for different time(0, 24, 48, 72 hours), the MTT method was used to detect growth proliferation of Hela cells in each group. Reverse transcription PCR was used to detect the changs of HIF-2α and VE-cadherin expressions at m RNA level in 100μmol/L Co Cl2 for 0, 24, 48, 72hours. Western blot was used to test the changs of HIF-2α and VE-cadherin expressions at protein level in 100μmol/L Co Cl2 for 0, 24, 48, 72 hours.Results: 1. By detecting results in 60 cases of cervical adenocarcinoma tissues and 18 cases of normal cervical tissues, we found that the positive expression rate of HIF-2α in cancer tissues was 56.67% and in normal cervical tissues was only 11.11%, the difference between which was statistically significant(P<0.05); and the positive expression rate of VE-cadherin in cancer tissues was 65.00% and in normal cervical tissues was merely 5.56%, the difference between which was statistically signifycant(P<0.05). The expression of HIF-2α and VE-cadherin was not related with pathological grading of cervical adenocarcinoma(P>0.05) and the age of patients but tumor size, lymph node metastasis and clinical staging of tumor(P<0.05). Otherwise, between HIF-2α and VE-cadherin there is a positive relationship, whose correlation coefficient was 0.632(P<0.05). 2.MTT results showed that the growth activity of human cervical cancer Hela cell line was enhanced when cells was cultured in 50μmol/L and 100μmol/L Co Cl2 for 24 hours(P<0.05),when 100μmol/L the cell line reached the maximum;And then,along with hypoxia prolonged,cell viability was decreased compared with the control group(P<0.05).The proliferation was inhibited by higher concentration of Co Cl2(P<0.05).3. Reverse transcription PCR results showed that, compared with normoxic control group, under the action of the 100μmol/L Co Cl2, HIF-2α and VE-cadherin in other time groups wereincreased, and the increase of VE-cadherin reached a maximum value undert 48 hours of hypoxia, whereas HIF-2α is has been on an upward trend. There was a positive correlation between the expression of HIF-2α and VE-cadherin at m RNA levels.The correlation was 0.987, and P=0.013(P<0.05).4.Western blot results showed that, compared with normoxic control group, under hypoxia environment induced by 100μmol/L Co Cl2 for different time, Hela cells of HIF-2α and VE-cadherin expressions were increased, and the VE-cadherin expression reached the maximum amount for 48 hours in hypoxia, then decreased, while the expression of HIF-2α protein has been a rising trend. There was a positive correlation between the expression of HIF-2α and VE-cadherin at protein levels 0.869,P=0.031(P<0.05). Conclusion: 1. The expression of HIF-2α and VE-cadherin at protein levels may have some role in predicting to patients in tumor invasion and metastasis. 2. HIF-2α may be one of the VE-cadherin regulatory factors, and HIF-2α may be by means of inducing VE-cadherin protein expression to increase the invasion and metastasis opportunity of cervical adenocarcinoma. 3. Hypoxia could promote the proliferation of cervical carcinoma Hela cell line. When the concentration of Co Cl2 increased and the time prolonged, the proliferation of cervical carcinoma Hela cell line inhibited.4. HIF-2α may be by means of inducing VE-cadherin protein expression to reduce the inhibitory effect on the proliferation of Hela cells under hypoxia. 5. HIF-2α inhibitors and antiangiogenic agengts may have potential basis and clinical study value onthe treatment of cervical adenocarcinoma, and can make a supplement for the kinds of novel anticancer drugs and chemotherapy drugs in cervical adenocarcinoma.