Abstract
Objective To construct the lentiviral vector of milk fat globule-epidermal growth factor 8 (MFG-E8) short hairpin RNA (shRNA), and stably transfected the HCC1973 cell line to detect its interference efficiency and influence in breast cancer cell HCC1973. Methods The knockout gene fragment was synthesized and inserted into the shRNA lentiviral vector PLKO.1 with a cherry protein reporter. The constructed recombinant plasmids were transfected into 293T cells respectively. The lentiviruses were transfected into HCC1973 cells and stably transfected PLKO.1-MFG-E8 shRNA recombinant lentiviruses were obtained. The expression of MFG-E8 was detected by real-time PCR and Western blotting. The cell proliferation was measured by methyl thiazol tetrazolium (MTT) assay and Flow cytometry was used to detect the cell cycle and apoptosis, Transwell was used to detect the invasion and migration of cells. Statisticalanalysis of data using the statistical product and service solutions 17.0 software. Results The constructed vector of PLKO.1-MFG-E8 shRNA significantly reduced the expression of MFG-E8 gene and protein in breast cancer cell HCC1973, the lentivirus titer was 3×108 PFU/ml. The proliferation of breast cancer cells was significantly suppressed (t=4.426, P<0.05), G0/G1 phase cells increased significantly with a decrease of cells in S phase (t=2.264, P<0.05), the ability of cell invasion and migration was significantly reduced (t=4.802, P<0.05), after the lentiviral infection. Conclusion The expression of MFG-E8 gene and protein by shRNA interference can inhibit the malignant biological behavior of breast cancer cells effectively, which indicates that MFG-E8 is involved in the invasion, metastasis and apoptosis of breast cancer during the process of breast carcinogenesis. Key words: Milk fat globule-epidermal growth factor 8 short hairpin RNA; Cell cycle; Cell proliferation; Cell poptosis; Invasion and metastasis
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