IASPP And CDKL1Expression And The Effect Of RNA Interfering On Tumour Cells Biological Activity

Objective: This study aimed to evaluate the expressions of iASPP (Inhibitor ofapoptosis stimulatory protein phosphatase) and CDKL1(Cyclin dependent kinase like1) in cancer tissues and to investigate their roles in the development and malignantprogression cancer.Methods: Real-time PCR (RT-PCR) and Western blot separately were used toanalyze the mRNA expression level and protein level of iASPP in fifty-six pairs ofovarian tissues including ovarian cancer tissues and the matched adjacent tissues.Receiver operating characteristic (ROC) analysis was performed to further evaluatethe clinical value of iASPP mRNA expression for the prediction of ovarian cancer. Inhuman ovarian cell line OVCA420, iASPP gene expression was silenced with aniASPP-siRNA interference method. After transfected, CCK-8assay was employed totest cells proliferation activity and the cells apoptosis was determined with Hoechst33342.In the work, the CDKL1mRNA level in fresh biopsy tissues from186breastcancer patients, with98benign tissues as negative control. CDKL1protein in30paraffin-embedded tissues from primary breast cancer patients were detected byreal-time RT-PCR assay and immunohistochemical staining, respectively. Roles ofCDKL1in cell growth were analyzed with CDKL1shRNA inhibitor-transfected cells.Results: iASPP is over-expressed in human ovarian cancer tissues when comparedwith the matched adjacent tissues (P=0.001). The expression of iASPP was significa-ntly decreased in the cells of OVCA420after the transfection of siRNA (P<0.05).Down-regulation of iASPP can be effectively inhibited cell proliferation (P<0.05) and significantly induced apoptosis.CDKL1was over-expressed in breast cancer patients and had a positive detectionefficiency of77%(144/186), which showed statistically significant differencecompared with ER, PR, P53, VEGF and E-cad (P<0.05). Inhibiting CDKL1functionwith shRNA, MCF-7cells exhibited obviously accumulation at G2/M phase andincreased sensitivity to cell cycle chemotherapeutic drugs.Conclusion: The over-expression of iASPP and CDKL1in cancer tissues suggestedthat they play respectively important roles in the development cancer, and the mRNAexpression level could be helpful for early diagnosis of cancer. The behavior afterRNA interference indicated that iASPP and CDKL1could be a potential target forcancer therapy.