Shrna Downregulation Of CacyBP/SIP Gene Expression On The Proliferation Of Human Osteosarcoma Cells

Objective: Osteosarcoma is a common primary malignant bone tumor.The incidence of osteosarcoma is increasing gradually.The comprehensive therapy is the regular treatment which is consisted of chemotherapy,surgery and chemotherapy,with lower five-year survival rate,About 60%-70%.We need to make a breakthrough in the treatment of osteosarcoma,in order to provide experimental basis and theoretical basis for chemotherapy and gene therapy for osteosarcoma patients.In this experiment,we tried to adjust shRNA to reduce CacyBP(calcyclin binding protein/Siah-1-interacting protein,CacyBP /SIP)gene and to observe its effect on SaOS-2 cell strain of human osteosarcoma.This article also discuss the possibility of silencing CacyBP /SIP gene therapy for osteosarcoma.Methods:1 U2 OS,HOS,MG-63 and Saos-2 a total of 4 kinds of human osteosarcoma cell strains were growing to detect CacyBP/SIP mRNAs expression abundance in different strains with the methods of real-time quantitative PCR(Application of real time quantitative polymerase chain reaction).2 We applied shRNA lentivirus infection of target cells,and divided the Saos-2 cells strain into 2 groups: the shCACYBP group(sh CACYBP lentivirus infection group)and shCtrl group(negative control virus infection group).3 Real-time quantitative PCR was used to detect the expression of CacyBP/SIP mRNA in the cells of the 2 groups.And investigate the effect of knock down of CacyBP/SIP gene mRNA in Saos-2 cells strain.4 The expression of CacyBP/SIP protein in the cells of the 2 groups was detected by Western blotting,and the before and after expression of CacyBP/SIP protein in Saos-2 cells strain was compared.5 The colony forming ability of the 2 groups in the cell culture plate was detected by the colony forming assay,and the tumorigenic ability of shRNA lentivirus infection on human osteosarcoma cell line Saos-2 was compared.6 MTT assay was used to detect the cell viability of the 2 groups of cells,and the effect of proliferation of human osteosarcoma cell line Saos-2 was compared.Results:1 Average ΔCts of Real-time quantitative PCR detection of CacyBP/SIP gene in U2 OS,HOS,MG-63 and Saos-2 were 4.71 ± 0.103,3.56 ± 0.075,3.80 ± 0.070,4.22 ± 0.110.CacyBP/SIP gene in 4 osteosarcoma cell lines were higher expression abundance.2 The expression of CacyBP/SIP mRNA in shCACYBP group and shCtrl group was detected by Real-time quantitative PCR Methods.The shCACYBP group(0.269 ±.0.014)was lower than that of shCtrl group(1.001 ± 0.060),with statistical significance(P<0.05).3 The expression of CacyBP/SIP protein in shCACYBP group and shCtrl group was detected by Western blotting,and the expression of CacyBP/SIP protein was decreased in sh CACYBP group.4 The number of cell clones in shCACYBP group and shCtrl group was 45 ± 8 and 112 ± 6,respectively.The number of clones in shCACYBP group was lower than that in shCtrl group,with statistical significance(P<0.05).5 MTT assay,shCACYBP group and shCtrl group with the change of time,the 2 groups in the eliasa absorption rate for the wavelength of 490 nm and light absorption rate were increased with the change of time.Compared with the shCtrl group,shCACYBP group Cell viability decreased,with statistical significance(P<0.05).Conclusions:1 The high abundance expression was detected in CacyBP/SIP in U2 OS,HOS,MG-63 and Saos-2 cells strains.2 The CacyBP / SIP mRNA level and protein expression of human osteosarcoma cell line Saos-2 can be decreased after shRNA lentivirus infection.3 Down-regulation of CacyBP / SIP expression in human osteosarcoma cell line SaOS-2 can inhibit the proliferation of SaOS-2 cells.4 CacyBP / SIP can promote tumor growth and proliferation in human osteosarcoma cell line SaOS-2.CacyBP / SIP is a Carcinogenic gene.