Expression Of LncRNA CCAT2,miR-205-5p,PTEN In Blood Of Breast Cancer Patients And Effect On Lymph Node Metastasis

Objective:To investigate the expression levels of CCAT2、miR-205-5p and PTEN in the blood of breast cancer patients,then to explore the effect and prediction of CCAT2,miR-205-5p and PTEN on lymph node metastasis of breast cancer patients,so as to provide new potential markers and ideas for detecting breast cancer metastasis.Methods:Go analysis and KEGG pathway enrichment analysis were performed for the downstream target factors of miR-205-5p,and PPI network interaction analysis plots among the target factors were also used.Patients with breast cancer diagnosed and untreated at the initial case in the Department of breast surgery,Southwest Medical University Hospital,were enrolled and divided into metastatic and para metastatic groups based on the presence or absence of lymph node metastasis.Forty-three patients were included in each group,and fresh blood samples from the patients were collected.The relative expression of CCAT2,miR-205-5p and PTEN in whole blood was determined by q RT-PCR and the m RNA levels were determined using the 2^-ΔΔct.Multivariate logistic regression was used to analyze the risk factors for metastasis in breast cancer patients.Sensitivity and specificity as well as ROC curves were used to evaluate the predictive effect of CCAT2,miR-205-5p and PTEN,and Spearman rank correlation was used to analyze the linear association between CCAT2,miR-205-5p and PTEN.Results:（1）Results of the raw signal analysis:the downstream target factors of miR-205-5p were searched in seven databases and their intersection was taken,resulting in 22 downstream target factors of miR-205-5p.Go analysis and KEGG pathway analysis of its 22 target factors,as well as PPI network interaction analysis map among the 22 target factors,were finally combined with the corresponding literature to finally determine the downstream target of miR-205-5p as PTEN.（2）Expression results of CCAT2,miR-205-5p and PTEN in the blood of breast cancer patients the expression levels of CCAT2 and PTEN were higher in the metastasis group than in the non-metastasis group,and the differences were statistically significant(Z_CCAT2=4.703,P<0.001;Z_PTEN=4.332,P<0.001);The expression of miR-205-5p in the metastasis group was lower than that in the non-metastasis group,and the difference was statistically significant（Z=3.692,P<0.001）.（3）Results from multivariate logistic analysis of the effects of CCAT2,miR-205-5p and PTEN on lymph node metastasis in breast cancer patients:CCAT2 and PTEN were risk factors for metastasis in breast cancer patients(OR_CCAT2=2.531,95%CI:1.130～5.587,OR_PTEN=4.718,95%CI:1.702～13.078),and miR-205-5p was a protective factor for metastasis in breast cancer patients（OR=0.361,95%CI:0.145～0.897）.（4）Analysis of the predictive effect of CCAT2,miR-205-5p and PTEN on breast cancer lymph node metastasis showed that when CCAT2>1.14,the sensitivity was 60%,specificity 91%,area under the curve 0.77,and concordance test（Kappa=0.512,P<0.001）;With miR-205-5p>1.07,it has a sensitivity of 65%,specificity of 81%,area under the curve（AUC）of 0.74 and concordance test（Kappa=0.465,P<0.001）for breast cancer metastasis prediction;When PTEN>1.22,the sensitivity for breast cancer metastasis prediction was 63%,the specificity was 84%,the area under the curve was 0.79,and the concordance test（Kappa=0.465,P<0.001）.The combined prediction of two indicators,CCAT2+miR-205-5p,CCAT2+PTEN,and PTEN+miR-205-5p,showed a sensitivity of 93%,88%,79%,a specificity of 72%,62%,77%,and an area under the curve of 0.85,0.83,and 0.80,respectively.Applying the three index combined prediction,the sensitivity was 93%,specificity 74%,and area under the curve 0.87;after dividing the three factors into high and low expression according to the cut-off value,multivariate logistic regression analysis showed that breast cancer patients with high CCAT2expression had a 7.665-fold higher risk of developing metastasis than those with low expression（OR=7.665,95%CI:1.684～34.891）;breast cancer patients with high PTEN expression were 7.861 times more likely to develop metastasis than those with low expression（OR=7.861,95%CI:2.000～30.901）,breast cancer patients with high expression of miR-205-5p had 0.186-fold higher risk of developing metastasis than those with low expression（OR=0.186,95%CI:0.045～0.768）.（5）Linear association analysis between CCAT2,miR-205-5p,and PTEN:there was a negative relationship between CCAT2 and miR-205-5p（r_s=-0.263,P<0.05）,a negative relationship between miR-205-5p and PTEN（r_s=-0.223,P<0.05）,and a positive relationship between CCAT2 and PTEN（r_s=0.278,P<0.001）.Conclusion:（1）The expression of CCAT2 and PTEN was higher in the breast cancer metastasis group than in the non-metastasis group,and the expression of miR-205-5p was lower in the metastasis group than in the non-metastasis group;（2）High CCAT2 and PTEN expression are risk factors for breast cancer metastasis,and high miR-205-5p expression is a protective factor for lymph node metastasis in breast cancer patients.（3）Furthermore,it had some predictive value for breast cancer lymph node metastasis when CCAT2>1.14,PTEN>1.22,and miR-205-5p>1.07,and the combination of the three indexes predicted breast cancer lymph node metastasis best.