| Breast cancer is one of the most common malignant tumors of women.The morbidity of breast cancer has been the first of female malignant tumor in our country.It has important impact on the health of women.Breast cancer includes many multiple subtypes,the Invasive carcinoma of no specific type(ductal NST)is the most common type,its factors are lack of regular structure,organized characteristics and different pathological morph.Mucinous carcinoma(MC)is a special type of breast cancer,which is characterized by the low degree of differentiation,the obvious cellular atypia,and the diffuse infiltrating lymphocytes.Tumor infiltrating lymphocytes(TILs)are mainly heterogeneous lymphocytes that exist in tumers,including T cells,B cells and natural killer cells(NK).T cells are lymphoid stem cell that derive from bone marrow and differentiate in the thymus,with the role of killing target cells directly.Its surface marker is CD3.According to the surface markers,T cells could be divided into CD4+T cells and CD8+T cells.CD4+T cells are called helper T cells(Th),and CD8+T cells are called cytotoxic T cells(Tc).B cells are derived from the lymphatic stem cells in bone marrow,their surface marker is CD20.B cells perform humoral immune function.NK cells also differentiate from the bone marrow stem cells,which can kill tumor cells directly.And their surface marker is CD57.Interleukin 2(IL-2)is secreted by T cells especially Th cells when stimulated by antigen or mitogen.It can stimulate the proliferation and differentiation of T cells and induce cytotoxic effect,it also has a certain role in promoting B cells growth and differentiation.Lymphotoxin α(LT α),is secreted by lymphocytes when activated by antigen or mitogen and it can also be secreted under the growth of some tumors.LT α can participate in the regulation of immune response,induce growth and differentiation of a variety of cells and have cytotoxic effect on a variety of tumor cells.In this study,immunohistochemistry was used to analyse the TILs,IL2 and LTα in two breast cancers’ patients with ductal NST and MC,and then analysed the effect of TILs,IL2 and LTα on clinical stage and prognosis of the patients with breast cancer.Objective: To investigate the effects of TILs,IL-2 and LTα on clinical stage and prognosis of the two breast cancers’ patients with MC and ductal NST.Methods:119 breast cancer patients in the Fourth Hospital of Hebei Medical University from 2007 to 2010 were selected as the study subjects,which included 76 cases of ductal NST and 43 cases of MC.Then HRP method of immunohistochemistry was used to analyze the distribution and expression of TILs,IL-2 and LTα in the two breast cancers.TILs were expressed in the cell membrane,and the judgement of immunohistochemistry was divided into two types according to the difference of the tumor stroma and parenchyma.The rate positive expression of the stromal compartment was to selected 10 visual of 200× to count TILs’ area percentage,namely stromal infiltrating lymphocytes area / stromal area,then we concluded the average density of stromal infiltrating lymphocytes.The positive expression rate of the intratumoral compartment: to selected 10 visual of 400× to count the number of TILs infiltrating in intratumoral.LTα and IL-2 are both expressed in the cytoplasm or cell membrane of lymphocytes and tumor cells.Randomly selecting visions,and according to the proportion of positive cells and staining intensity to analyze.Achromatic color was marked of 0 point,pale yellow was marked of 1 point,pale brown was marked of 2 points,and brown was marked of 3 points.Proportion of positive cells: the negative cell was 0,the positive cells < 10% were 1 point,11%~50% were 2 points,51%~75% was 3 points,and >75% was 4 points.The percentage of positive cells and the product of staining intensity were used as result: 0 points was for the negative(-),1~4 were divided into weak positive(+),5~8 were moderately positive(++),9~12 were strong positive(+++).Results:1 Main clinicopathological featuresThe age of onset of ductal NST was 29-77 years’ old,the median age was 51.43 years old.The age of onset of MC was 28-77 years’ old,the median age was 50.51 years old.There was no significant difference between the two groups(P=0.669).The diameter of tumor tissue in ductal NST group was 1cm-5cm,the average diameter was 2.11 cm.The diameter of tumor tissue in MC group was 0.5cm-4cm,the average diameter was 1.89 cm.The difference was not statistically significant(P=0.278).The lymph node metastasis rate of ductal NST group was 57.89%(44/76).The lymph node metastasis rate of MC group was 25.58%(11/43).The lymph node metastasis rate of ductal NST group was higher than that of MC group,and the difference was statistically significant(P=0.001).The positive rate of ductal in NST group was 35.52%(27/76),and the MC group was 13.95%(6/43),and the difference was statistically significant between NST ductal group and MC group(P=0.012).The clinical stage of NST ductal group stage I were 20 cases,stage II were 45 cases,stage III / IV were 11 cases;in group MC,stage I were 17 cases,stage II were 25 cases,stage III / IV were 1 cases.The clinical stage of ductal NST group was higher than that of MC group,the difference was statistically significant(P=0.036).In MC group,ER,PR and HER2 were negative,molecular classification were Basal-like type,so there was no correlation analysis.The CD3,CD4,CD8,CD20,CD57,IL-2 and LTα in ductal NST group did not have relationship with ER,PR,HER2,Ki-67 and molecular classification,the difference was not statistically significant(P>0.05).2 Experimental results of immunohistochemistry with HRP method2.1 Tumor infiltrating T lymphocytesIn 76 cases of ductal NST,the median of stromal cell density of CD3+T cells was 5%,and the median of tumor infiltrating cells were 2/HPF.In 43 cases of MC,the median of stromal cell density of CD3+T cells was 40%,and the median of tumor infiltrating cells were 15/HPF.In interstitium and parenchyma,the CD3+T lymphocytes in MC group were both higher than those in ductal NST group,and the difference was statistically significant(P=0.000,P=0.000).In 76 cases of ductal NST,the median of stromal cell density of CD4+T cells was 2%,and the median of tumor infiltrating cells were 1/HPF.In 43 cases of MC,the median of stromal cell density of CD4+T cells was 10%,and the median of tumor infiltrating cells were 5/HPF.In interstitium and parenchyma,the CD4+T lymphocytes in MC group were both higher than those in ductal NST group,and the difference was statistically significant(P=0.000,P=0.000).In 76 cases of ductal NST,the median of stromal cell density of CD8+T cells was 3%,and the median of tumor infiltrating cells were 1/HPF.In 43 cases of MC,the median of stromal cell density of CD8+T cells was 30%,and the median of tumor infiltrating cells were 10/HPF.In interstitium and parenchyma,the CD8+T lymphocytes in MC group were both higher than those in ductal NST group,and the difference was statistically significant(P=0.000,P=0.000).In MC group,in interstitium and parenchyma the Tc lymphocytes were higher than Th,and the difference was statistically significant(P=0.000,P=0.004).In ductal NST group,in interstitium and parenchyma there was no significant difference between Tc and Th,which was not statistically significant(P=0.511,P=0.307).2.2 Tumor infiltrating B lymphocytesIn 76 cases of ductal NST,the median of stromal cell density of CD20 cells was 3%,and the median of tumor infiltrating cells were 0.In 43 cases of MC,the median of stromal cell density of CD8+T cells was 15%,and the median of tumor infiltrating cells were 2 / HPF.In interstitium and parenchyma,the CD20 lymphocytes in MC group were both higher than those in ductal NST group,and the difference was statistically significant(P=0.000,P=0.001).2.3 Tumor infiltrating NK lymphocytesIn 76 cases of ductal NST,the median of stromal cell density of CD57 cells was 0,and the median of tumor infiltrating cells were 0.In 43 cases of MC,the median of stromal cell density of CD8+T cells was 1%,and the median of tumor infiltrating cells were 1 / HPF.In interstitium and parenchyma,the CD8+T lymphocytes in MC group were both higher than those in ductal NST group,and the difference was statistically significant(P=0.000,P=0.000).In the MC group,the expression of stromal CD3 was higher than that of CD20,the difference was statistically significant(P=0.000);the expression of stromal CD20 was higher than that of CD57,and the difference was statistically significant(P=0.000).That means the stromal T cells were the most,B cells were in the middle,and NK cells were the least.And the expression of intratumoral CD3 was higher than that of CD20,the difference was statistically significant(P=0.000);CD20 and CD57 were not significantly different,and the difference was not statistically significant(P=0.391).That means the intratumoral T cells were the most,B and NK cells had no difference.In the ductal NST group,the expression of stromal CD3 and CD20 were not significantly different,and the difference was not statistically significant(P=0.082);the expression of stromal CD3 and CD20 were higher than CD57,and the difference was statistically significant(P=0.000).There was no significant difference in the expression of stromal T and B cells,and NK cells were lower than that of the formers.And the expression of intratumoral CD3 was higher than that of CD20,the difference was statistically significant(P=0.000);the expression of intratumoral CD20 was higher than that of CD57,and the difference was statistically significant(P=0.002).That means the intratumoral T cells were the most,B cells were in the middle,and NK cells were the lowest.2.4 IL-2In 76 ductal NST cases,24 cases were +,43 cases were ++ and 9 cases were +++.In 43 MC cases,5 cases were +,17 cases were ++ and 21 cases were +++.The expression of IL-2 in MC group was higher than that in ductal NST group,and the difference was statistically significant(P=0.000).2.5 LTαIn 76 ductal NST cases,11 cases were +,38 cases were ++ and 27 cases were +++.In 43 MC cases,4 cases were +,11 cases were ++ and 28 cases were +++.The expression of LTα in MC group was higher than that in ductal NST group,and the difference was statistically significant(P=0.007).3 The effect of CD3,CD4,CD8,CD20,CD57,IL-2 and LTα on the clinical stage of the two breast cancers3.1 The effects of single factor CD3,CD4,CD8,CD20,CD57,IL-2 and LTα on the clinical stage of the two breast cancersCD3,CD8 and CD20 had effect on clinical T stage of MC group,the difference was statistically significant(P=0.026,P=0.048,P=0.044).CD4,CD57,IL-2 and LTα did not have obvious effect on clinical T stage of MC group,there was no statistically significant difference(P>0.05).CD3,CD8 and LTα had effect on clinical N stage of MC group,the difference was statistically significant(P=0.021,P=0.027,P=0.035).CD4,CD20,CD57 and IL-2 did not have obvious effect on clinical N stage of MC group,there was no statistically significant difference(P>0.05).CD3,CD4,CD8,CD20,CD57,IL-2 and LTα did not have obvious effect on either clinical T or N stage of ductal NST group,there was no statistically significant difference(P>0.05).3.2 The effect of multi factors CD3,CD4,CD8,CD20,CD57,IL-2 and LTα on the clinical stage of the two breast cancersCD8 had effect on clinical T stage of MC group,the difference was statistically significant(P=0.017).CD8 can be used as a independent factor which had influence on clinical T stage of MC.CD8 had effect on clinical N stage of MC group,the difference was statistically significant(P=0.034).CD8 can be used as a independent factor which had influence on clinical N stage of MC.CD3,CD4,CD20,CD57,IL-2 and LTα did not have obvious effect on either clinical T or N stage of MC group,there was no statistically significant difference(P>0.05).CD3,CD4,CD8,CD20,CD57,IL-2 and LTα did not have obvious effect on either clinical T or N stage of ductal NST group,there was no statistically significant difference(P>0.05).4 The effect of CD3,CD4,CD8,CD20,CD57,IL-2 and LTα on survival rate of the two breast cancers4.1 The effect of single factor CD3,CD4,CD8,CD20,CD57,IL-2 and LTα on survival rate of the two breast cancersCD8 had effect on the survival rate of MC group,the difference was statistically significant(P=0.024).CD3,CD4,CD20,CD57,IL-2 and LTα did not have obvious effect on the survival rate of MC group,there was no statistically significant difference(P>0.05).CD3,CD4,CD8,CD20,CD57,IL-2 and LTα did not have obvious effect on the survival rate of MC group,there was no statistically significant difference(P>0.05).4.2 The effect of multi factors CD3,CD4,CD8,CD20,CD57,IL-2 and LTα on survival rate of the two breast cancersThe 5-year survival rate of MC group(93.0%)was higher than that of ductal NST group(78.9%),the difference was statistically significant(P=0.048).The high expression of CD8 was positively correlated with the prognosis of the MC group,the difference was statistically significant(P=0.013).CD3,CD4,CD20,CD57,IL-2 and LTα had no significant effect on the prognosis of the MC group,the difference was not statistically significant(P>0.05).CD3,CD4,CD8,CD20,CD57,IL-2 and LTα had no significant effect on the prognosis of the ductal NST group,the difference was not statistically significant(P>0.05).Conclusions:1 The total T cells,Th cells,Tc cells,B cells,NK cells in interstitium and parenchyma of MC group were more than those of ductal NST group.It is suggested that the clinical stage and prognosis of MC group was better than ductal NST group may have relationship with the infiltrating T,B and NK cells,and especially T cells and Tc cells may play an important role.2 The expression of IL-2 and LTα in MC group was higher than ductal NST group.It is suggested that the clinical stage and prognosis of MC group was better than ductal NST group may have relationship with IL-2 and LTα.3 CD8 can be used as an independent prognostic factor which has an effect on the clinical stage and prognosis of MC group.4 CD3,CD4,CD8,CD20,CD57,IL-2 and LTα had no significant effect on 5-year survival rate of the ductal NST group... |
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