| microRNA(micro-RNA,miRNA),a class of 22~24 necleotide non-coding RNAs,modulate gene expression through interacting with binding to the target messenger RNA.Accumulating reports have shown that miRNA are frequently dysregulated in cancer and play a key role in the pathology process of breast cancer and its inoccurrence,development,invasion and metastasis,which indicates that miRNAs may promise as miRNA-based biological markers for cancer diagnosis and prognostication.Emerging evidence has demonstrated that miRNA involve in regulating lots of physiological process and disease,including cell proliferation,apoptosis,stem cell differentiation,inoccurrence and metastasis and so on.miRNA has been considered as the most promising as biological molecules for the prevention and treatment of tumor and other diseases.Breast Cancer(BC)is the most common cancer type and the second cause of cancer-related death among women.Death caused by metastasis in breast cancer is still the leading cause of death,and it is difficult to date has not yet been solved.Using new the knowledge of life science to explore miRNA of molecular mechanism of breast cancer regarding regulation of development,improving the accuracy and specificity of diagnosing early breast cancer,diagnosing and treating early,adopting treatment measures to treat breast cancer before the breast cancer cells metastasize,which have crucial significance to improve the cure rate of breast cancer and save life for breast cancer patients.In our study,we first use literature analysis and cancer cell lines in vitro,combine with clinical samples data analysis,then screened and confirmed miRNA that regulate in breast cancer metastasis and invasion,screened non-invasive miRNA biomarkers in plasma of patients for noninvasive early diagnosis of breast cancer.Using our own design microRNA microarray and performing qRT-PCR to analyze abnormal miRNA expression in plasma of breast cancer,we designed miRNA microarray with high specificity,accuracy,efficient and convenient,celerity.In our study,we previously have collected individuals plasma samples from40 healthy control object and 54 Breast Cancer.Curculating microRNA were identified by performing global microRNA microarray on in plasma of 54 Breast Cancer and 40 healthy control objects.Then we found that members of circulating miR-17-92a cluster family,miR-17-5p,miR-19a,miR-19b,miR-92a and circulating miR-27a have been subsequently confirmed as significantly overexpressed in breast plasma samples.In terms of these miRNA,we explored the correlation between miRNA and pathophysiology and clinical features of breast cancer.Materials and Methods1.Clinical dataFrom January 2012 to January 2016,the 54 breast cancer patients and matched 40healthy control subjects were recruited from Department of thyroidea of Shanghai Dong fang Hospital.2.Plasma specimen collection and preservationWhole blood(5ml)before treatment were draw into EDTA-containing tubes.Informed consent was obtained from the breast cancer patients.All blood were processed in 4 hours,collected plasma was aliquoted and stored in-80?.3.microRNA chip screeningPlasma from the 54 breast cancer patients and matched 40 healthy control subjects performed microRNA chip by ABI 7900.The expression level was normalized to 5s rRNA and was represented by utilizing the 2~-ddctddct methord.ddH2O was used as negative control.4.Statistical analysisSPSS 24.0 software was used for statistical analysis.In our study,Mann-Whitney(-Wilcoxon)test was used for comparisons of 2 independent groups.ROC curves was performed to evaluated the diagnostic value of plasma miRNA for differentiating breast cancer patients from healthy control subjects.Results1.miRNA in plasma of breast screening and the diagnostic value of breast cancerplasma miRNAIn our study,microRNA microarray and qRT-PCR were used to detected miRNA expression in plasma of 54 breast cancer patients and 40 controls.Comparing expression of miRNA in plasma samples from the control subjects,we found miR-17-5p,miR-19a,miR-19b,miR-92a,miR-27a was were in plasma smaples from breast cancer patients significantly up-regulated.Their p value between plasma samples of breast cancer and the control subjects were statistically significant.Area under the receiver operating characteristic curve(AUC)was used to valued diagnostic accuracy of plasma miRNA,the result were 0.707,0.663,0.706,0.687,0.672.2.The differential miRNAs of breast cancer plasma and collection with breast cancermetastasisComparing the controls,miR-17-5p(Z=-2.247,P=0.025),miR-19b(Z=-2..718,P=0.007),miR-92a(Z=-3.718,P=0.000)and other circulating miR-27a(Z=-1.969,P=0.049)was up-regulated in plasma from patients with non-metastatic breast cancer.Comparing the controls,miR-17-5p(Z=-2.190,P=0.029),miR-19b(Z=-2.580,P=0.010)was up-regulated in patients with metastatic breast cancer.Comparing the metastatic breast cancer,level of miR-92a(Z=-2.218,P=0.033)in non-metastatic breast cancer was more higher.3.The differential miRNAs of breast cancer plasma and collection with invasivebreast cancerComparing the controls,we found Circulating miR-17-5p(Z=-2.838,P=0.005),miR-19a(Z=-2.522,P=0.012),miR-19b(Z=-2.135,P=0.033),miR-92a(Z=-2.645,P=0.008)and other circulating miR-27a(Z=-2.595,P=0.009)was all up-regulated in plasma from non-invasive breast cancer.Comparing the controls,circulating miR-17-5p(Z=-2.589,P=0.010),miR-19a(Z=-3.901,P=0.000),miR-19b(Z=-3.314,P=0.001),miR-92a(Z=-2.853,P=0.004)and other circulating miR-27a(Z=-2.436,P=0.015)was all up-regulated in plasma of invasive breast cancer.4.The differential miRNAs of breast cancer plasma and collection with clinical stagesIn view of Breast cancer clinical stages,miR-17-5p,miR-19a,miR-19b,miR-92a,miR-27a were correlation with T1,T2.In addition,miR-19b and miR-92a was associated with T3-4 stage of breast cancer.Conclusions1.The plasma level of miR-17-92a cluster family,including miR-17-5p,miR-19a,miR-19b,miR-92a and other circulating miR-27a was significantly up-regulation inbreast cancer patients compared with healthy control.2.Exploring the correlation between miRNA and breast cancer metastasis,we foundthat miR-17-5p,miR-19b,miR-92a and other circulating miR-27a was up-regulatedin plasma from patients with metastatic breast cancer and patients withnon-metastatic breast cancer.Among these dysregulated miRNAs,level of miR-92awas more higher in plasma of metastatic breast cancer than non-metastatic breastcancer.3.Exploring the relationship between miRNA and invasive/non-invasive breast cancer,we found Circulating miR-17-5p,miR-19a,miR-19b,miR-92a,and othercirculating miR-27a was all up-regulated between plasma from invasive/non-invasive breast cancer.4.Investigating the correlation between miRNA and pathological staging of breastcancer,we found that the circulating miR-17-5p,miR-19a,miR-19b,miR-92a andother circulating miR-27a in plasma of the breast cancer patients correlated with T1and T2 stage of breast cancer.In addition,miR-19b and miR-92a was associatedwith T3-4 stage of breast cancer. |
PDF Full Text Request