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Abstract
To investigate the relationship between the expression of p21WAF1/ClP1protein and p53 status and the possible role of the two proteins in hepatocellular carcinomas (HCCs), we examined the expression of p21WAF1/CIP1and p53 immunohistochemically in 81 tumours from 65 patients with hepatocellular carcinoma. p21WAF1/CIP1protein was absent from 59 of 81 tumours (72.8%), and altered p53 expression was found in 43 (53.1%). p21WAF1/CIP1expression was significantly associated with p53 status (P= 0.0008); 38 of 59 tumours lacking p21WAF1/CIP1protein were accompanied by altered p53 expression. Further analyses showed that p21WAF1/CIP1expression was inversely correlated with p53 expression in hepatitis C virus (HCV)-related HCCs, but not in HBV-related hepatocellular carcinomas and hepatocellular carcinomas without viral infection. All 11 tumours with intrahepatic metastasis showed altered p21WAF1/CIP1or p53 expression. In contrast, no intrahepatic metastasis was found in any of the 17 tumours without abnormal expression of either of the two proteins. These results suggest that: (1) different modes of p21WAF1/CIP1regulation are involved in HCCs differing in their hepatitis viral infection status, and p21WAF1/CIP1expression appears to be predominantly related to altered p53 in HCV-related HCCs; (2) disruption of the p53–p21WAF1/CIP1cell- cycle-regulating pathway may contribute to malignant progression of HCC. © 2000 Cancer Research Campaign
Highlights
Background liver diseaseChronic hepatitis 15/18 (83.3%) 9/18 (50.0%) Liver cirrhosis 44/63 (69.8%) 34/63 (54.0%) Tumour size
Cell cycle progression is governed by several checkpoints, which are regulated by a family of protein kinases, the cyclin-dependent kinases (CDKs) and the cyclins (Hunter and Pines, 1994)
We previously investigated p21WAF1/CIP1 mRNA expression by reverse-transcriptase polymerase chain reaction (RT-PCR) and p53 mutational status by PCR single-strand conformation polymorphism (SSCP) and direct DNA sequencing in hepatocellular carcinomas (HCCs)
Summary
After brief washing with distilled water, tissue sections were processed in 10 mM citrate buffer (pH 6.0) and heated to 120°C in an autoclave for 10 min for antigen retrieval (Hui et al, 1999a, 1999b, 2000; Li et al, 2000). Slides were allowed to cool at room temperature for 20 min and rinsed with phosphate-buffered saline (PBS). To inhibit non-specific binding activity, slides were incubated with blocking serum at room temperature for 30 min. The sections were incubated with biotinylated anti-mouse immunoglobulins (Vector Laboratories, Inc., Burlingame, CA, USA) for 30 min, and avidin–biotin complex (Vector Laboratories) for 30 min at room temperature, with washing in PBS before each incubation.
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