Effect Of Pirarubicin Collaborate With Hepacam Gene On Proliferation And Cell Cycle In Renal Cell Carcinoma

Part one: Hematuria detection and mRNA expressions of cyclinB1andhepaCAM in RCCObjective: To explore hematuria in urine specimens and mRNA levelsof cyclinB1and hepaCAM in renal cell carcinoma(RCC).Methods Dry chemistry method and urinary sediment microscopyanalysis were used to detect hematuria in50urine specimens. RT-qPCR wasused to test the mRNA levels of cyclinB1and hepaCAM in the32pairedrenal specimens. Analyze the relationships between the results and theclinical parameters.Results In50cases of RCC patients,15cases(30%) pay attention tomacroscopic hematuria,29cases(58%) were positive in dry chemistryoccult blood test, and27cases (54%)were positive in microscope detection.In32cases of RCC tissues, CyclinB1mRNA was down-regulated in26cases(81.3%), hepaCAM mRNA was down-regulated in28cases(85.7%).The expression of CyclinB1and hepaCAM in RCC tissues were significantly lower than that in adjacent tissues(P＜0.05). There was norelationship between cyclinB1,hepaCAM mRNA expressions and thepatient′s age, gender，tumor size，subtype and nuclear grade.Conclusion The low discovery rate of macroscopic hematuria in RCCpatients may be one of the reason that cause treatment delay, dry chemistrymethod and urinary sediment microscopy analysis can increase thedetection rate of hematuria, and then contribute to the early detection ofRCC. CyclinB1and hepaCAM are significantly decreased in RCC, but therelationship between cyclinB1and hepaCAM still lack reliable evidence.This is a basic premise for the study of postoperative treatment on RCC. Part tow: pirarubicin collaborate with hepaCAM gene to furtherdown-regulate cyclinB1in renal cell carcinomaObjective:To investigate the effects of pirarubicin（THP） cooperatedwith hepaCAM gene on renal cell carcinoma.Methods After treated with THP,786-0cells were infected withadenovirus pAdH5-hepaCAM or pAdH5. MTT assay was used to observethe inhibition rates of cells in different groups. FCM was used to analyze the changes of cell cycle. RT-qPCR was used to detect the mRNA level ofhepaCAM and cyclinB1.Western blot and immunofluorescence were used todetect the protein expressions of hepaCAM and cyclinB1.Results MTT assay showed inhibition rate in THP/pAdH5-hepaCAMgroup was markedly increased as compared with that in THP/pAdH5groupand pAdH5-hepaCAM group (P＜0.01); In FCM assay, remarkable G2/Mphase arrest was seen in THP/pAdH5-hepaCAM group; RT-qPCR show themRNA level of cyclinB1was significant down-regulated inTHP/pAdH5-hepaCAM group(P＜0.01). Western blot andimmunofluorescence showed the protein level of cyclinB1wasdown-regulated in THP/pAdH5-hepaCAM group.Conclusion When gene transfer used after THP treatment, remarkabledecrease of cyclinB1can cause G2/M phase arrest and then cause significantgrowth inhibition. We presumed that the collaboration of THP andhepaCAM can reduce the recurrence of RCC after operation.