MLPH Regulates The Biological Behavios Of Triple Negative Breast Cancer And Its Mechanism

Objective:The occurrence of invasion and metastasis of breast cancer is affected by cell movement.Potential biological targets related to cell movement are screened by comparing the sequencing results with the databases.The impact of the target on the biological behavior of breast cancer is further demonstrated in combination with clinical pathological specimens and biological experiments,and its potential molecular mechanism is preliminarily discussed.Methods:Combined with sequencing results and database analysis,screening potential biological targets of breast cancer.Immunohistochemical staining was used to verify the relationship between its expression and clinicopathological features in breast cancer,and immunofluorescence was used to observe the target gene location.Western Bolt and q-PCR were used to select the expression of target genes in breast cancer cell lines.The lentivirus transfected was used to knockdown or overexpress target gene,the transfection efficiencies were detected by Western Bolt and q-PCR.The plate cloning,CCK-8 assay and Transwell chamber assay were used to test the ability of proliferation,migration and invasion in breast cancer cells.Immunofluorescence assay was used to observe the localization changes of target gene.Western Bolt was used to detect the expression of epithelial-mesenchymal transformation-related proteins by the changes of target gene expression.Result:Bioinformatics analysis showed that MLPH was specifically expressed in breast cancer and correlated with the expression of AR.Immunohistochemistry showed that the relationship between MLPH and clinicopathological features was consistent with that of AR.MLPH was mainly expressed in the cytoplasm,while AR was mainly expressed in the nucleus.There was a correlation between the expression of MLPH and AR.The expression of MLPH in various breast cancer cell lines was detected by Western Bolt and q-PCR,and MDA-MB-231 was selected as the experimental cell line to construct the cell model.Plate cloning and CCK8 experiments showed that the expression of MLPH was related to cell proliferation:the proliferation decreased after MLPH knockdown,and increased after MLPH over-expression.Transwell chamber experiment suggested that the expression of MLPH was related to the migration and invasion ability of cells:the migration and invasion ability were reduced after MLPH knockdown,and the migration and invasion ability were enhanced after MLPH over-expression.Western Bolt results and immunofluorescence assay showed that the expression of MLPH could affect the protein level of AR and its localization in MDA-MB-231 breast cancer cells.Meanwhile,Western Bolt results showed that expressions of E-cadherin,N-cadherin and Vimentin related to epithelial-mesenchymal transformation were affected by MLPH:The expression of E-cadherin increased and the expression of N-cadherin and Vimentin decreased after the expression of MLPH was decreased,after over-expression of MLPH,the expression of E-cadherin decreased,and the expression of N-cadherin and Vimentin increased.Conclusions:MLPH can affect the ability of proliferation,migration and invasion in breast cancer cells,mainly plays a similar role as a tumor promoter in MDA-MB-231 cells.The mechanism may be closely related to MLPH regulating EMT process or affecting AR expression.