Effect Of Proliferation,migration And Invasion Of Tri-negative Breast Cancer And Clinical Correlation Analysis

Objective(s):Triple Negative Breast Cancer(TNBC)is a special type of Breast Cancer.Its malignancy and risk of recurrence and metastasis are higher than other Breast Cancer subtypes,and the prognosis of patients is relatively worse.Several studies have shown that insulin-like Growth Factor 2(IGF2)is associated with the occurrence,progression,metastasis and prognosis of a variety of cancers.In this study,the clinical correlation between IGF2 and triple negative breast cancer and its effect on the proliferation,migration and invasion of triple negative breast cancer were investigated by using triple negative breast cancer tissue samples and triple negative breast cancer cell lines.Methods:1.Sample collection:Fifty-two cases of invasive breast cancer(tri-negative type)diagnosed by postoperative pathology without preoperative endocrine therapy,preoperative radiotherapy,neoadjuvant chemotherapy or targeted therapy who visited the Department of Breast Surgery of the First Affiliated Hospital of Kunming Medical University(the main hospital)from January 2015 to September 2018 were collected.Postoperative pathology confirmed 12 cases of invasive breast cancer(hormone receptor positive type)2.The expression of IGF2 in triple negative breast cancer tissues and hormone receptor positive breast cancer tissues was detected by Immunohistochemistry(IHC)of all paraffin specimens collected.3.To analyze the expression of IGF2 in paraffin tissue of triple negative breast cancer and hormone receptor positive breast cancer tissue,and further analyze the statistical differences between the expression of IGF2 and the clinical characteristics of TNBC cancer patients,such as age,Ki-67 expression,TNM stage,axillary lymph node metastasis,recurrence and metastasis,and abortion history.4.According to the immunohistochemical results of IGF2 expression,expressed in the patients with TNBC-,+,++as low expression group,+++as a high group,analysis of IGF2 expression and 3 years of two groups of patients with TNBC disease-free survival time(diseases-free survival,DFS),And Overall survival(OS)at 3 years.Cox regression was used to analyze prognostic factors.5.The effects of IGF2 at gradient concentrations on the proliferation,migration and invasion of breast cancer cells were detected:(1)CCK8 cell proliferation experiment testing different concentrations IGF2 after processing,the three negative breast cancer cell lines and hormone receptor-positive breast cancer cell line proliferation ability.(2)Cell scratch test was used to detect the migration ability of three kinds of breast cancer cells in IGF2-treated group and control group,the migration distance between the control group and the experimental group was measured and the differences in migration rates between the groups were analyzed.(3)Transwell test was used to detect the migration and invasion ability of the three kinds of cells in the control group and the IGF2-treated group,and the number of membrane penetrating cells in each group was counted and the differences between the groups were analyzed.Results:1.Expression of IGF2 in two types of breast cancer tissues:Immunohistochemical results show that the samples in this study,IGF2 in three negative breast cancer tissue and hormone receptor-positive breast cancer tissues were expressed,IGF2 strong positive in three negative breast cancer tissue type is higher than the proportion of hormone receptor-positive breast cancer tissue,immunohistochemical IGF2 expression differences between two groups has no statistical significance(P>0.05)2.Correlation analysis results of IGF2 expression and clinical indicators in TNBC tissueIn 52 cases of TNBC patients,the analysis of the relationship between IGF2 expression and clinical indicators showed that there was statistical significance between Ki-67 value,recurrence and metastasis,history of abortion and IGF2 expression of patients(P<0.05),while there was no statistical significance between age,TNM stage and IGF2 expression of TNBC patients(P>0.05)3.Survival analysis and prognostic factors analysis of IGF2 high expression group and low expression group in TNBC:The relationship between 3-year DFS and IGF2 expression in the high and low IGF2 expression groups in TNBC patients was analyzed by drawing a survival curve.The relationship between 3-year DFS and OS in the high expression group and the low expression group of IGF2 in TNBC patients was analyzed by drawing the survival curve,and it was found that there was statistically significant difference in 3-year DFS and IGF2 expression in the two groups(P<0.05),while there was no statistically significant difference in 3-year OS and IGF2 expression in the two groups(P>0.05)Cox univariate analysis showed that IGF2 was correlated with 3-year DFS in TNBC patients(P<0.05),and age was correlated with 3-year OS in TNBC patients(P<0.05).Multivariate analysis showed no statistical significance(P>0.05).4.Effects of IGF2 at gradient concentrations on proliferation,migration and invasion of three kinds of breast cancer cells:(1)In the CCK8 cell proliferation experiments,three kinds of breast cancer cells of difference between control group and IGF2 treatment groups have statistical significance(P<0.05),the OD value of the difference between cells of different concentration groups has no statistical significance(P>0.05).(2)In the cell scratch experiments,IGF2 treatment group of MDA-MB-231 cells and MDA-MB-468 cells’ ability to migrate and the control group have statistical significance difference between(48 h:24 h,P>0.05,P<0.05)in(24 h:P<0.05,48 h,P<0.05).There was no significant difference in the migration ability between MCF-7 cells and the control group(24h:P>0.05,48h:P>0.05).(3)In the Transwell migration experiments,IGF2 after processing of MDA-MB-231,MDA-MB-468 cells and MCF-7 cell migration ability and the differences between the control group was statistically significant(P<0.01,P<0.05,P<0.05).(4)In Transwell invasion experiment,IGF2 MDA-MB-231 cells after processing,its attack ability is better than the control group(P<0.01),the difference was statistically significant.The invasion ability of MDA-MB-468 cells treated by IGF2 was statistically significant different from that of the control group(P<0.05),while the invasion ability of MCF-7 cells was not statistically significant different(P>0.05)Conclusions:1.The proportion of IGF2 strongly positive in triple negative breast cancer tissues was higher than that in hormone receptor positive breast cancer tissues,suggesting that IGF2 may be related to the malignant degree of tumor.2.The expression intensity of 20IGF2 in TNBC patients with recurrent metastasis,ki-67 20%,and abortion history was different from that of TNBC patients without recurrent metastasis,ki-67<20%,and without abortion history,suggesting that IGF2 may have certain predictive effect on recurrence and metastasis of patients with triple-negative breast cancer.3.Compared with the low expression group,the 3-year DFS difference of TNBC patients in the high expression group of 20IGF2 was statistically significant,while the 3-year OS difference between the two groups was not statistically significant,and the multifactor analysis was not statistically significant,suggesting that IGF2 may have certain research value for the 3-year DFS of TNBC patients.However,age and IGF2 were not independent influencing factors for the prognosis of TNBC patients.4.IGF2 to three negative breast cancer cells and hormone receptor-positive breast cancer cell proliferation has a certain role in promoting.5.IGF2 may promote the migration and invasion of triple negative breast cancer cells,but it still needs to be further verified.