Study On Hedgehog Signaling Pathway Molecules In Breast Cancer And Breast Cancer Stem Cells

Objective:(1)To investigate the related gene expression of Hedgehog signaling pathway and its significance of breast cancer stem cell with primary breast cancer and mathed lymph node metastasis.(2)To investigate the relationship between the related protein espression of Hedgehog signaling pathyway and ER,PR.Methods:30cases of fresh breast invasive ductal carcinoma and matched metastasis lymph nodes were made into single cell suspensions by mechanical separation, then breast cancer stem cells were separated from them by immunomagnetic sorting. The related gene expressions of Hedgehog signaling pathway of breast cancer stem cells were detected by Real time PCR. The related protein expressions of Hedgehog signaling pathway and ER,PR were detected by immunohistochemistry in a series of57human invasive breast cancer.Results:(1)The expression of Shh mRNA was28.96±4.58in primary breast cancer stem cells, and the expression of Shh mRNA relative to β-actin was20.59±7.12in primary breast cancer stem cells; The expression of Shh mRNA was14.1±6.52in matched lymph node metastases cancer stem cells,and the expression of Shh mRNA relative to β-actin was25.19±7.36in matched lymph node metastases cancer stem-cells. The expression of Shh mRNA had no differences in primary breast cancer stem cells and matched lymph node metastases cancer stem cells(P>0.05).The expression of Ptch mRNA was24.59±1.12in primary breast cancer stem cells, and the expression of Ptch mRNA relative to β-actin was20.23±3.33in primary breast cancer stem cells; The expression of Ptch mRNA was25.09±1.48in matched lymph node metastases cancer stem cells,and the expression of Ptch mRNA relative to P-actin was19.29±4.36in matched lymph node metastases cancer stem cells. The expression of Ptch mRNA in primary breast cancer stem cells was higher than it in matched lymph node metastases cancer stem cells(t=-1.825, P<0.05,2-△△CT=2.69).The expression of Smo mRNA was31.12±3.21in primary breast cancer stem cells, and the expression of Smo mRNA relative to β-actin was19.71±4.42in primary breast cancer stem cells; The expression of Smo mRNA was29.75±3.18in matched lymph node metastases cancer stem cells,and the expression of Smo mRNA relative to β-actin was18.65±5.34in matched lymph node metastases cancer stem cells. The expression of Smo mRNA in primary breast cancer stem cells was lower than it in matched lymph node metastases cancer stem cells(t=0.354,P<0.05,2-△△CT=0.81).The expression of Gli-1mRNA was33.13±3.97in primary breast cancer stem cells, and the expression of Gli-1mRNA relative to β-actin was18.78±5.51in primary breast cancer stem cells; The expression of Gli-1mRNA was31.09±2.28in matched lymph node metastases cancer stem cells,and the expression of Gli-1mRNA relative to β-actin was17.49±5.44in matched lymph node metastases cancer stem cells. The expression of Gli-1mRNA in primary breast cancer stem cells was lower than it in matched lymph node metastases cancer stem cells(t=0.580,P<0.05,2-△△CT=0.58).(2)The positive rates of Shh,Smo and Gli-1in normal breast ductal epithelial tissues were31.4%、28.6%and20%,while the positive rates of Shh, Smo and Gli-1in breast cancer were87.7%、89.5%and89.5%.The difference was statistically significant between normal brest ductal epithelial tissues and breast cancer(P<0.01)(3) The positive rates of Shh, Smo and Gli-1in breast cancer were91.2%,91.2%and73.7%.The positive rates of ER in breast cancer was54.4%. The expression of ER had no significantly correlated with Shh and Gli-1(P>0.05),While it was significantly correlated with Smo(rp=-0.261, P=0.05).The positive rates of PR in breast cancer was35.1%. The expression of PR had no significantly correlated with Shh,Smo and Gli-1(P>0.05).Conclusion:(1)Comparing with the primary breast cancer stem cells,the Hedgehog signaling pathway had higher activated condition in the cancer stem cells of matched lymph node metastases. The Hedgehog signaling pathway of breast cancer stem cells may play an important role in proliferation and differentiation, invasion and metastasis of breast cancer.So we concluded that the breast cancer stem cells which had higher activated condition of Hedgehog signaling pathway in primary breast cancer could be easier to metastasize in lymph node.(2)The Hedgehog signaling pathway related proteins show higher expression in breast cancer, and Smo had significantly correlated with ER. The higher expression of Smo in the Hedgehog signaling pathway may control the expression of ER.