The data demonstrated although no change in the expression of Cdk1 was seen, that treatment of cells with PLAB or colchicine increased the expression of cyclin B1. Taken together, the data suggested that PLAB induced cell cycle arrest in U87 glioblastoma cells at M phase but perhaps not at G2 phase. p53 is among the most powerful tumor Hedgehog inhibitor suppressor genes in human cancers. Paid down the influence of PLAB and PFT, a p53 inhibitor, because U87 glioblastoma cells express wild type p53, we desired to observe the expression of p53 in PLABtreated U87 cells using Western blot. We discovered that PLAB markedly increased the expression of p53 in U87 cells in a dose-dependent fashion. Because Bax is one of the critical downstreammediators of p53 signalling, we observed the possible changes in the expression of Bax.
A heightened expression of Bax was present in PLAB addressed U87 cells. Besides the induction of Bax, p53 service has been proven to inhibit the expression of anti-apoptotic protein Bcl 2 and our Western blot analysis unmasked the same. To help define the apoptosis process, we tested the expression of cytochrome c and caspase Inguinal canal 3 in U87 glioblastoma cells. The data showed that PLAB increased the expression of cytochrome c in cytosol and cleaved the caspase 3 into 17 kDa and 12 kDa proteins. To further confirm the contribution of caspase 3 in PLABinduced apoptosis in U87 glioblastoma cells, we noticed the expression of caspase 3 substrate, PARP using Western blot. Figure 7 shows the cleavage of PARP in to 85 kDa protein. These results obviously show that PLAB causes caspase 3 dependent apoptosis in U87 glioblastoma cells.
As shown in Figure 4, the overall caspase inhibitor, z VADfmk did not inhibit the apoptotic effect of PLAB completely. This suggests that some caspase independent apoptotic pathway can also be involved. Apoptosis inducing factor has been reported to induce caspase independent apoptosis Ganetespib by straight inducing DNA fragmentation. We wished to check whether AIF is involved with PLAB induced apoptosis in cells. We examined the effect of PLAB on AIF nuclear translocation usingWestern mark. PLAB therapy increased the expression of AIF in nucleus dose dependently, as shown in Figure 7. Hepatotoxicity and nephrotoxicity are the major negative effects of cancer chemotherapeutic agents. Consequently, we investigated the effect of PLAB on liver and kidneys using Kunming mice. The cytotoxic effect of PLAB was assessed by measuring the changes in body-weight, blood biochemistry and histopathology of liver and kidneys in comparison with get a grip on group. No apparent change in body weight of mice in treatment group has been observed when comparing to get a handle on group.
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