ABSTRACT
Objective To observe the PML protein expression of hepatocellar carcinoma tissue and cells lines and As 2 O3 regulate its expression .Methods Immunohistochemistry was used to examine the PML protein expression of hepatocellar carcinoma tissue . Western blot analysis were used to observe PML protein expression of hepatocellar carcinoma tissue of 12 cases ,5 hepatocellar car-cinoma cell lines ,such as HuH7 ,HepG2 ,Hep3B ,SMMC-7721 ,MHC97H .Western blot analysis was used to detected the PML pro-tein expression of these hepatocellar carcinoma cell lines after 72-96 h treated with 0 .25 μg/mL of As2 O3 .Results Immunohisch-enmical staining showed that the PML protein was expressed in both cytoplasm and nucleus ,did not well-distributed in hepatocellar carcinoma cells .There was no significant differences of PML protein expressed among differently differentiated stages of hepatocel-lar carcinoma cells .Western blot analysis found that hepatocellar carcinoma tissues of 12 cases with hepatocellar carcinoma ex-pressed PML protein ,and there was significant difference of PML protein expressed among 12 cases suffer with hepatocellar carci-noma .hepatocellar carcinoma cell lines ,such as HuH7 ,HepG2 ,Hep3B ,SMMC-7721 and MHC97H all expressed PML protein ,and there was little difference of PML protein expressed among hepatocellar carcinoma cell lines .The PML protein expression of HuH7 ,HepG2 ,Hep3B ,SMMC-7721 and MHC97H cell after 72-96 h treated with 0 .25 μg/mL of As2O3 significant decreased . Conclusion Hepatocellar carcinoma tissue and cells may express PML protein ,and As2 O3 may regulate this protein expression as well .PML protein may be the target molecule of As2 O3 treating HCC .
ABSTRACT
Background and purpose:Recent researches have shown that Secreted Protien Acidic and Rich in Cysteine (SPARC) was closely related to tumor genesis, tumor progression and tumor metastasis. SPARC was highly expressed in malignant melanoma, glioma, meningioma, bladder cancer, lung cancer and prostate cancer, etc. In this study we investigated SPARC expression in hepatocellar carcinoma (HCC) and its signifi cance. Methods:RT-PCR was used to detect SPARC mRNA expression in cancer tissue samples and their adjacent liver tissue samples from 62 patients with hepatocellar carcinoma and 30 normal liver tissue samples, respectively. And the differential protein expression of SPARC between these groups was analyzed by immunohistochemistry (IHC). Results:SPARC mRNA was highly expressed in HCC(14.0?3.6) and in the adjacent liver tissue (6.8?1.8); compared with low expression of 2.7?0.9 in normal liver tissue, there were signifi cant differences among the three groups (p=0.000). SPARC positively stained was found in 54 of 62 patients with HCC and 4 of 30 normal liver tissue, there was significant difference between these two groups (P=0.000). SPARC immunohistochemical score was 21.5?4.8 in the carcinoma group; 11.3?3.6 in paracarcinoma group and 5.7?1.8 in the normal group, there were also significant differences among the three groups (P=0.000). The expression of SPARC protein was significantly upregulated with the progress of Enmondson pathological classification. There was obviously differences between Ⅰvs Ⅱ(P=0.029), and Ⅱ vs Ⅲ Ⅳ(P=0.008). There was more SPARC expression in the patients with metastasis of HCC (26/27, 96.3%) than that without metastasis(23/35, 65.7%)(P=0.004). Conclusion:SPARC mRNA expression and its protein were related to HCC histological differentiation and metastatic lymph node; SPARC is helpful to clinical evaluation of HCC.
ABSTRACT
Purpose:To investigate the effects of tumor suppressor gene p53 on the activity of telomerase/hTERT in human HCC-9204 cell line.Methods:Lipofection-mediated gene transfection method was used to transfect wild-type p53(wt-p53) gene into HCC 9204 cell which expresses telomerse /hTERT and carries mutant-type p53 gene. The expression of p53 was confirmed by Western blot. Both the telomerase activity and hTERT mRNA expression were detected by PCR-ELISA,TRAP-silver staining,in-situ hybridization and RT-PCR methods. The apoptotic appearance was examined by FCM.Results:Higher telomerase activity and hTERT mRNA level were detected in HCC-9204,and they were markedly inhibited after transfection with wt-p53. Meanwhile,decreasing level of bcl-2 protein and appearance of apoptosis were also shown in the transfected cells.Conclusions:Over expression of the exogenous wt-p53 gene does suppress both telomrease activity and hTERT mRNA expression in HCC cell line. There is a p53-dependent regulatory pathway for activation and expression of telomerase/hTERT in HCC.