| Backgroung:Bladder cancer is one of the most common malignant tumors of the genitourinary system.The incidence rate and mortality rate in China are increasing year by year.The pathogenesis of bladder cancer is very complex.The abnormal expression or function of many proteins and the change of many signal pathways may affect the occurrence and development of bladder cancer.The main reason for the limitation of CT/MRI in the prediction of positive lymph nodes in bladder cancer is that it is unable to locate small volume and micrometastasis lymph nodes,the early diagnosis technology is lack of specificity and sensitivity,and the high postoperative recurrence rate is still the main problem in this fieldsTn antigen is closely related to tumor proliferation,differentiation and metastasis.Studies have found that sTn antigen can promote the activity,infiltration and immune escape of tumor cells,so it is closely related to lymph node metastasis.Differential expression of sTn may be related to P13K/Akt/mTOR pathway and EMT.However,the expression of GSK3β,mTOR and sTn in bladder cancer tissues,adjacent tissues,lymph node tissues,normal tissues and blood,and the correlation between GSK3β,mTORC and sTn and the relationship between sTn and lymph node metastasis are not clear.Objectives:Study the expression of GSK3β,mTOR,sTn in cancer tissues,adjacent tissues,lymph node tissues,normal tissues,and blood1.Whether there is a correlation between the expression of sTn,GSK3β and mTOR in the sample;2.sTn,GSK3β and mTOR are differentially expressed in normal bladder tissue,non-muscle-invasive bladder cancer tissue(NMIBC)and muscle-invasive bladder cancer tissue(MIBC)3.sTn,GSK3P,and mTOR in cancer tissue and lymph tissue have differential expressions for lymph node metastasis4.Early screening and postoperative detection of lymph node metastasis of bladder cancer by differential expression of STN,GSK3β and mTOR genes in bloodMethods:1.(1)four patients with NMIBC and MIBC were selected respectively(inclusion criteria:patients did not receive chemotherapy,radiotherapy,immunotherapy and other treatments before operation),and one normal control(patients with bladder angioma).(2)Six patients with lymphnode metastasis,two with lymphnode metastasis and one normal control(bladder angioma)were selected.2.Collect cancer tissue,paracancerous tissue,lymph node tissue,normal tissue and blood tissue and put them into-80°refrigerator for preservation.The gene expression of GSK3β,mtorc and STN in cancer tissue,paracancer tissue,lymph node tissue,normal tissue and blood were detected by PCR.Expression of immunohistochemistry in cancer tissue,paracancerous tissue,lymph node tissue and normal tissue.Results:1.Q-PCR test results(1).In lymph group and blood group,the gene expression of GSK3β and mTOR was positively correlated(P<0.01);in cancer group,the gene expression of GSK3βand mTOR was not statistically significant(P=0.160)(P>0.05);in adjacent group,the gene expression of GSK3β and mTOR was not statistically significant(P=0.276)(P>0.05).(2).Nine cases were divided into normal group(No.1),NMIBC group(No.2,3,4,5)and MIBC group(No.6,7,8,9).GSK3β and mTOR in cancer tissue were compared with those in the above three groups.For mTOR protein gene expression,NMIBC group was higher than normal group,the difference was statistically significant(P<0.01),MIBC group was higher than NMIBC group,the difference was statistically significant(P<0.01);For GSK3β protein gene expression,NMIBC group was higher than normal group,the difference was statistically significant(P<0.01),MIBC group was higher than NMIBC group,the difference was statistically significant(P<0.01).(3).Nine cases were divided into normal group(No.1),non-lymphatic metastasis group(No.7,8)and lymphatic metastasis group(No.6,9).Regarding the expression of mTOR protein gene in cancer tissues,the non-metastatic group had higher bladder tissue than the normal group,and the difference was statistically significant(P<0.01).The lymphatic metastasis group was higher than the non-metastatic group,and the difference was statistically significant(P<0.01);for GSK3β protein gene expression in cancer tissues,the bladder tissues in the non-lymphatic metastasis group were higher than those in the normal group,the difference was statistically significant(P<0.01),and the lymphatic metastasis group was higher than the non-lymphatic metastasis,the difference was significant Statistical significance(P<0.01).The 9 cases were divided into two groups:the non metastasis group(no.7,8)and the metastasis group(no.6,9).For the expression of mTOR protein gene in lymphoid tissue,the difference was statistically significant(P<0.05).For the expression of GSK3β protein gene in lymph tissue,the difference was statistically significant(P<0.01).(4)Nine cases were divided into normal group(No.1),non metastasis group(No.7,No.8)and metastasis group(No.6,No.9).For the expression of mTOR protein gene in the blood,the difference was statistically significant(P<0.01)between the lymph node metastasis group and the normal group(P<0.01).For the expression of GSK3β protein gene in the blood,the difference was statistically significant(P<0.01)between the lymph node metastasis group and the normal group(P<0.01)2.Immunohistochemical test results(1)In the lymph group and the cancer group,sTn was positively correlated with GSK3β and mTOR(p<0.01),while in the paracancer group,sTn had no correlation with GSK3β and mTOR(p>0.05).(2)Divide 9 cases into normal group(No.1),NMIBC group(No.2,3,4,5)and MIBC group(No.6,7,8,9),cancer tissues sTn,GSK3β and mTOR will be The above three groups are compared in pairs.The expression of sTn protein in cancer tissues was higher in the NMIBC group than in the normal group,and the difference was statistically significant(P<0.05).The MIBC group was higher than the NMIBC group High,the difference was statistically significant(P<0.01);mTOR protein expression in cancer tissues,NMIBC group was higher than the normal group,the difference was statistically significant(P<0.01),MIBC group was higher than NMIBC group,the difference was statistically significant(P<0.05);GSK3β protein expression in cancer tissue,NMIBC group was higher than normal group,the difference was statistically significant(P<0.01),the MIBC group was higher than the NMIBC group,the difference was statistically significant(P<0.05).(3)Group 9 cases into normal group(No.1),non-lymphatic metastasis group(No.7,8)and lymphatic metastasis group(No.6,9).The expression of sTn protein in cancer tissues was higher in the non-metastatic group than in the normal group,and the difference was statistically significant(P<0.01).The lymphatic metastasis group was higher than the non-metastatic group,and the difference was statistically significant(P<0.05);The expression of mTOR protein in cancer tissues was higher in the lymphatic metastasis group than in the normal group,and the difference was statistically significant(P<0.05).The lymphatic metastasis group was higher than the lymphatic metastasis group,and the difference was statistically significant(P<0.05);The expression of GSK3β protein in cancer tissues was higher in the non-metastatic group than in the normal group,and the difference was statistically significant(P<0.05).The lymphatic metastasis group was higher than the non-metastatic group,and the difference was statistically significant Significance(P<0.01).The 9 cases were divided into two groups:the non metastasis group(no.7,8)and the metastasis group(no.6,9).The expression of sTn protein in lymph tissue was higher in lymph node metastasis group than that in non metastasis group(P<0.01);and the expression of mTOR protein in lymph tissue was higher in the group of lymph node metastasis than that in the group of lymph node non metastasis(P<0.05).The expression of GSK3β protein in lymphoid tissue was higher in lymph node metastasis group than that in non metastasis group(P<0.05).Conclusions:1.In lymphoid and blood tissues,the gene expression of GSK3β and mTOR is positively correlated;in cancer and paracancer,the gene expression of STN,GSK3βand mTOR is not correlated;in cancer,lymphoid and blood tissues,the gene expression of sTn,GSK3β and mTOR is positively correlated;in paracancer,the gene expression of sTn,GSK3β and mTOR is not correlated.It is possible that sTn can activate P13K/Akt/mTOR signal pathway by combining with Gal-3,and mTOR expression is increased,further up regulating GSK3β expression.2.The gene expression of GSK3β and mtorc in normal bladder,NMIBC cancer and MIBC cancer increased in turn,and the protein expression of sTn,GSK3 β and mTOR in normal bladder,NMIBC cancer and MIBC cancer increased in turn,suggesting that the detection of the expression of sTn,GSK3 β and mTOR in cancer may be helpful to the stage of bladder cancer.3.The gene expression of GSK3 β and mTOR in cancer and lymphoid tissue was lower than that in lymph node metastasis group,and the protein expression of sTn,GSK3 β and mTOR in cancer and lymphoid tissue was lower than that in lymph node metastasis group,suggesting that the expression of sTn,GSK3 βand mTOR in cancer and lymphoid tissue may have some reference to whether the bladder cancer patients have lymph node metastasis Leading meaning.4.The gene expression of GSK3 β and mTOR in the blood was lower in the group without lymph node metastasis than that in the group with lymph node metastasis.There was a positive correlation between the expression of sTn,GSK3 βand mTOR protein in cancer,lymph node and blood tissue,suggesting that the early screening and prognosis monitoring of the lymph node metastasis of cystis cancer might be achieved through the differential expression of sTn,GSK3 β and mTOR in the blood. |
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