| Objective: Bladder cancer is the most common malignant tumor in the urinary system,and it is also one of the ten most common tumors in the whole body,and it ranks the first in the incidence of urogenital cancer in our country.And found in recent years the study of bladder cancer,bladder cancer cell surface high expression of apoptosis ligand 1(B7-H1,also known as PD-L1)in the process of its development play a vital role,so in B7-H1/PD-1 way research also increasingly deeper,and on the way of immune targeted therapy has definite clinical effect;,however,accept the treatment of patients still have quite a number of the condition of the poor curative effect and even invalid,lead to cannot be treated by the effective treatment of bladder cancer,so studying the invalid bladder cancer patients treated in the body,what changes happened,will provide new breakthrough for the treatment of bladder cancer.Known the life activity of tumor cells is dependent on the energy metabolism,as is an extremely important role in the immune escape for its survival is inseparable from the sugar metabolism of energy supply,and tumor cells adopt the method of aerobic glycolysis energy has been recognized by everyone,so this study by blocking apoptosis ligand 1(B7-H1)expression,to explore the changes of bladder cancer sugar metabolism,cellular immune escapes in bladder tumors and the tumor was revealed the relationship between glucose metabolism,and for those who accept B7-H1 / PD-1 way immunotherapy of invalid patients,to provide new ideas for the treatment of bladder cancer.Method:When the cultured bladder cancer cells(T24 cells)were blocked by b7-h1 blockers(MIH1 blockers),they were divided into two groups: the experimental group and the control group,and the following experiments were conducted with the control experiment.Fluorescent tags 2-deoxidation of the glucose(2-NBDG)as two groups of cells,optical probe training testing training after 0.5 h the fluorescence intensity of bladder cancer cells,to judge the bladder cancer cells after B7-H1 block,the change of the ability to absorb glucose;Then,under the condition of the same training(with the exception of B7-no other different H1 blocking)two groups of cells after 24 h training out of the culture medium,measured with lactic acid kit glycolysis metabolites of lactic acid content,to display the bladder cancer cells after blocking the ability difference of glycolysis;Finally,to extract protein,respectively,two groups of cells by Western blot test glycolytic key enzyme(HK-2,PKM-2,LDHA)and related signaling pathways(pten/pi3k/akt/m TOR)protein expression,reflect the specific impact the signal mechanism of glycolysis.Result:(1)the B7-H1 blockers block after 24 h,using flow cytometry instrument detection of bladder cancer cell surface expression of B7-H1,the control group was 9.179 ±0.1255,and B7-H1 blocking group,the expression of B7-H1 amount was 0.3957 ±0.02976,P < 0.05,the difference was statistically significant,shows that the experimental group after blocking B7-H1 expression quantity few;(2)the B7-H1 blocking group of bladder cancer cells and the control group 2-NBDG in bladder cancer cells,compared to the fluorescence intensity of B7-H1 blocking of the fluorescence intensity of OD value was 393.9 ± 1.803,and 469.7 ± 4.017 in the control group,a significant reduction in the fluorescence intensity can be seen that block group,showed that bladder cancer cell glucose uptake ability;(3)B7-H1 blocking group of cancer cells was 0.8254 ± 0.01993 in lactic acid content in the tendency for L,and the lactic acid content in the control group was 1.227 ± 0.01202 tendency for L,blocking group compared with control group,the metabolism of lactic acid content decreased obviously,reflects the blocking B7-H1 after aerobic glycolysis ability to bladder cancer cells;In the cancer cells of b7-h1 blocking group,the key enzyme of glycolysis was decreased by 40.01%,LDHA decreased by 49.56%,and pkm-2 decreased by 35.82%,both of which were statistically significant(p<0.05).Akt signaling pathways mediating glycolytic pathway proteins fell by 44.34%,the active form of p-Akt fell by 44.68%,and had statistical significance(p < 0.05),suggesting that the B7-H1 is by lowering glycolysis key enzymes such as the HK-2,LDHA,PKM-2 and pten/pi3k/AKT/m TOR protein synthesis is reduced,to block the glycolytic pathway bladder cancer cells.Conclusion:By detecting the fluorescence intensity of 2-nbdg in bladder cancer cells(T24 cells),we found that B7H1 block could reduce the glucose uptake of bladder cancer cells.And through the metabolites of lactic acid,the metabolic HK-2 key enzymes,PKM-2,LDHA and Akt,p Akt analysis in the signal path,and then we get the preliminary conclusion,B7H1 by blocking an enzyme metabolism and protein synthesis in the signal path,make bladder cancer cell glycolysis ability to drop,this approach can provide new ideas for treatment of bladder cancer. |
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