Research Of ABCG2, P-gp And Lymphocyte Subsets In Breast Cancer

In recent years, the incidence of breast cancer is increasing year by year,surgery, radiotherapy, chemotherapy and hormone therapy are still the maintreatment methods. The chemotherapy, which occupies an important positionin the treatment, and anthracycline-based drug is one of the foundation ofbreast cancer chemotherapy drugs.In the study drug-resistant was directlyaffected breast cancer chemotherapy effect, which became the importantproblem about chemotherapy failed.P-glycoprotein（P-gp） which coded bydrug-resistant genes MDR1high expression was one of the main reasons thatgenerated multidrug resistance （MDR）,it was also the most extensive,the onlyconfirmed mechanisms of resistance thorough research and clinicalpractice.Breast cancer resistance protein（BCRP/ABCG2）is the latest discoverydrug-resistant protein, which has the same substrate with P-gp:anthracycline-based drugs.High expression of ABCG2and P-gp suggestsanthracycline-based drug resistance,avoid use such drugs when chemotherapycan get better curative effect, so discussing ABCG2, P-gp expression in breastcancer can not only help to understand their own drug resistance, but also havea guiding role.Including breast cancer, the development of tumor is complicated,that hasa close relationship with body’s immune function. Organisms antineoplasticimmune mainly divide into T lymphocyte-mediated cellular immune and Blymphocyte-mediated cellular immune. When cancer occurs, the body’santineoplastic immune will play a role, CD4is auxiliary T lymphocytes surfaceantigen, CD8is inhibitory T lymphocytes surface antigen, CD19present in theB lymphocytes surface antigen, CD56present in the NK cells surfaceantigen.Through testing T, B lymphocyte subsets and NK cells expression tokonw the immune status in the laboratory. Ahmed ’s research about early hematopoietic cells shows that, in vitro the ABCG2can make the originalprogenitor cells increasing,damage CD+19lymphocyte’s development,reduce thetransplant mice B lymphocytes quantity; in the body ABCG2can increase theproportion of the progenitor cells and the number of father cells. in earlyhematopoietic process, ABCG2as a regulatory protein plays an important role.So research the relationship of ABCG2and lymphocyte subsets in breastcancer have a certain innovation significance.This study used immunohistochemical method to detect ABCG2, P-gpexpression in breast cancer, and flow cytometric to detect blood lymphocytesubsets of breast cancer patients, combined with clinical material, to study theresistance status and immune function of patients breast cancer, then analyzedits relevance.The first part: ABCG2, P-gp expression in breast cancerPurpose:By detecting the expression of breast cancer drug resistance protein（ABCG2/BCRP） and P-glycoprotein （P-gp） in breast cancer tissue, we tried toreveal the relationship between the former factors and tumor biologicalcharacteristics.Methods:We investigated the distribution and expression of ABCG2, P-gp in85breast cancers and40mammary gland fiber adenoma tissues byimmunohistochemistry. And compared these former factors with clinicalpathological datas of breast cancer.Results:1ABCG2protein expression: ABCG2protein was mainly expressed inthe membranes and part of the cytoplasm of tumor cells,and a small quantityof in mammary gland fiber adenoma. Statistical analysis revealed thatABCG2-positive stained rate was significant higher in breast cancers （43.5%）than that in mammary gland fiber adenomas （25.0%）（P<0.05）.2P-gp protein expression: P-gp protein was mainly expressed in the membrane and part of the cytoplasm of tumor cells,and a small quantity of inmammary gland fiber adenoma. Statistical analysis revealed that P-gp-positivestained rate was significant higher in breast cancers （34.1%） than that in fiberadenomas （15.0%）（P<0.05）（P<0.05）.3ABCG2and breast cancer clinical pathology material relationshipStatistical analysis revealed that the expression of ABCG2in breastcancer had no significant relation with age, lymph node metastasis, AJCCstage （P＞0.05）, but it was correlated with the biggest diameter of the tumor（P<0.05）. Expression rate of ABCG2protein with biggest diameter greaterthan2cm （52.6%） was significant higher than that in lower （25.0%）（P<0.05）.4P-gp and breast cancer clinical pathology material relationshipStatistical analysis revealed that the expression of P-gp protein in breastcancer had no significant relation with age, tumor biggest diameter, AJCCstage （P＞0.05）, but it was correlated with lymph node metastasis （54.8%）was significant higher than that in without （22.2%）（P<0.05）.5There was a positive relationship between ABCG2-positive expressionand P-gp-positive expression in breast cancer （r=0.219, P<0.05）.Conclusion:1Increased expression of ABCG2and P-gp protein in breast cancerindicated that ABCG2and P-gp were closely correlated with clinical usesensitive chemotherapy drugs and in view of the two proteins’s treatment inbreast cancer.2The expression rates of ABCG2protein were correlated with thebiggest diameter which manifest that were closely correlated withcarcinogenesis in breast cancer. P-gp protein were correlated with lymphnode metastasis which have a certain value with clinical judgment to theprognosis of patients in breast cancer.3The close relationship of ABCG2and P-gp in breast cancer indicatedthat both of them promote each other, have the same mediation and the sameway direction. The second part: Lymphocyte subsets count of early breast cancer patients.Purpose:By testing the T B lymphocytes and the ratio of NK cells of early breastcancer patients, we tried to know the immune function and clinicalsignificance of breast cancer status.Methods:Four color fluorescent marker flow cytometry was used to detect theperipheral blood lymphocyte subsets CD+3cells, CD++4cells, CD8cells, B cells,a relative of NK cells count in20breast cancers and42healthy check-ups（control）, analyzed the results.Results:1T lymphocyte subsets count: the average of CD+3cells in early breastcancer and control group were76.44±6.07%,72.01±7.02%, There weresignificant differences between two groups （P<0.05）, CD4+/CD8+averageratio were1.99±1.16,1.42±0.89, There were significant differences betweentwo groups（P<0.05）.2B lymphocyte subsets and NK cell count: the average of B lymphocytesin early breast cancer and control group were9.39±3.51%，11.56±2.54%,There were significant differences between two groups （P<0.05）; NK cellswere7.80±4.42%，14.91±4.87%, There were significant differences betweentwo groups （P<0.05）.Conclusion:1Increased T lymphocyte subsets CD+3cells, CD₄⁺/CD₈⁺in early breastgroup indicated that the body’s immune function was disorder in early breastcancer, and immune cells was in active state.2Compared with healthy people B lymphocytes in early breast groupwas reduced, which manifest the humoral immune function, NK ratio and theability of killing tumor was decline in the early breast cancer.The third part:Lymphocyte subsets count of advanced breast cancer patientsPurpose: By testing the T B lymphocytes and the ratio of NK cells of advancedbreast cancer patients, we tried to know the immune function and clinicalsignificance of breast cancer status.Methods:Four color fluorescent marker flow cytometry was used to detect theperipheral blood lymphocyte subsets CD+3cells, CD₄⁺cells, CD₈⁺cells, B cells,a relative of NK cells count in56breast cancers and42healthy check-ups（control）, analyzed the results.Results:1T lymphocyte subsets count: the average of CD₄⁺cells in advancedbreast cancer and control group were34.32±11.25%，43.00±8.99%, Therewere significant differences between two groups （P<0.05）,CD4+/CD8+average ratio were1.21±0.71,1.42±0.89,There were significantdifferences between two groups（P<0.05）.2B lymphocyte subsets and NK cell count: the average of B lymphocytesin advanced breast cancer and control group were8.18±6.81%，11.56±2.54%,There were significant differences between two groups （P<0.05）; NK cellswere13.95±11.50%，14.91±4.87%, There were no significant differencesbetween two groups （P＞0.05）.Conclusion:1Increased T lymphocyte subsets of CD₈⁺cells,but CD₄⁺cell count,CD₄⁺/CD₈⁺ratio down. which manifest the body’s immune cells in decline inadvanced breast cancer.2Reduced B lymphocytes in advanced breast cancer indicated that thehumoral immune function drops, and more immune cells more early reactionimmune status; NK ratio was not statistically significant the advanced breastcancer.The fourth part:ABCG2, P-gp and lymphocyte subsets in Breast cancerPurpose:By testing the expression of ABCG2, P-gp and T B lymphocytes and the ratio of NK cells, we tried to reveal the relationship and clinical significancebetween the former factors.Methods:1We investigated the expression of20breast cancers byimmunohistochemistry;2Four color fluorescent marker flow cytometry was used to detecte Tlymphocytes, B lymphocytes and the ratio of NK cells of the same patients.Results:1ABCG2and lymphocyte subsets of the relationshipStatistical analysis revealed that the expression of ABCG2had nosignificant relation with CD₄⁺lymphocytes, CD₈⁺lymphocytes, CD++4/CD8ratioof NK cells （P>0.05）, but it was correlated with CD+3lymphocyte （r=0.600）and B lymphocyte （r=0.503,P<0.05）.2P-gp and lymphocyte subsets of the relationshipStatistical analysis revealed that the expression of P-gp had no significantrelation with CD+3lymphocytes,CD₄⁺lymphocytes,CD₈⁺lymphocytes,CD₄⁺/CD₈⁺ratio and B lymphocyte （P>0.05）, but it was correlated with theratio of NK cells （r=0.471,P<0.05）.Conclusion:1The close relationship of ABCG2with CD+3lymphocytes and Blymphocyte in breast cancer indicated that resistant breast cancer proteinexpression may relate wih the immune function in breast cancer patients.2The close relationship of P-gp and NK cells in breast cancer indicatedthat P-glycoprotein expression may be related to kill function in breast cancerpatients.but there was correlation with T lymphocytes and B lymphocytes.