Study On The Effection Of Hec1 In Regulating The Paclitaxel-sensitivity In The Ovarian Cancer Cells

Objective At present,drug resistance has become the major reason of failure for malignant neoplasm.Recent studies discovered that Hec1,a regulating gene in mitotic checkpoint,is higher expression in many neoplasms and is associalated evidently with the poor prognosis of the part of the tumor.Thus,we provides great inlight into the effection of Hec1 in changing the ovarian cancer cells'sensitivity to paclitaxel by RNAi technology. Methods 1)the difference of expression of Hec1 between normal ovarian and ovarian cancer was detected by immunohistochemisty. 2)the cells of A2780 and SKOV3 were transfected by control siRNA and Hec1 siRNA,the untransfected cells as the blank. 3)The expression of Hec1 mRNA and related protein level were evaluated with quantitative real-time RT-PCR ,western blot analysis and immunofluorescence, respectively. 4) Inhibition of cell proliferation was determined by MTT, and the induction of apoptosis and mitosis were examined through flow cytometry (FACS) , western blot and immunofluorescence. Results 1) the expression of Hec1 in ovarian cancer is much higher than in the normal ovarian. 2) the expression of Hec1 after transfected Hec1 siRNA is evidently reduced compared with control either in mRNA or in protein level.The same result was found though the immunofluorescence. 3) inhibiting the Hec1 can inhibit the cells'proliferation .At the same concentration of taxol ,the cell viability is evidently descreased. 4) inhibiting the Hec1 can increase the apoptosis and the mitotic arrest of the cells campared with the control cells, However, inhibiting the Hec1 alone after 48 hours didn't increase the cells in subG1, which represents the apoptosis. Conclusion the inhibition of Hec1 expression,which activate the mitotic checkpoint to induce the mitotic arrest,can result in chemosensitivity ability to paclitaxel in ovarian neoplasm. Objective To investigate the effection of MicroRNA-133(miR-133) expression on the invasive and migrative ability of Breast Cancer lowly metastatic cell line MCF-7 by regulation the expression of RHOA protein. Methods (1) MCF-7 cells were transfected with has-miR-133a inhibitor using Lipofectamine. MCF-7 Cells transfected with Negative control were cultured as control,untreated cells as blank.(2)the expression of miR-133 in transfected MCF-7 cells was detected by qRT-PCR (3) the effection of miR-133 on the migration and proliferation of transfected MCF-7 cells was detected by Transwell experiment and MTT. (4) The changes in skeleton of transfeced cells was observed by immunofluorescence. (5) The expression of RHOA protein regulated by miR-133 was detected by western blot. Results (1) the expression of miR-133 after transfected has-miR-133a inhibitor is evidently reduced compared with controls (2)Inhibiting miR-133 can enhance the migrative ability of MCF-7 cells,but no effection on proliferation. (3) The skeleton of cells transfected with has-miR-133a inhibitor was evidently changed compared with that of control,which represents the former has powerful ability of invasion and migration. (4) The level of RHOA protein in cells transfected with has-miR-133a inhibitor was significantly higher than that in two control cells detected by Western blot. Conclusion MiR-133 can suppress the invasion and migration of Breast Cancer cell line MCF-7 through regulating the expression of RHOA protein.