| Hepatocarcinoma is one of the most malignant human cancer, usually detected later and developing faster. Chemotherapy is a kind of measure that can effectively treat cancer. It mainly kill tumor cell through apoptosis. Apoptosis is a process of self destruction that requires the activation of a genetic programme that may lead to changes in cell morphology, DNA fragmentation, and protein cross linking. Less apoptosis than normal may lead to carcinogenesis. Recent study indicated that chemotherapeutic drugs, especially DNA-damage drugs, can activate NF-kB when they come into effect. NF-kB is a kind of transcription factor than exists extensively, act diversely and is related to immunity react apoptosis inflammation injury, etc. The experiment intents to explore the relation between the expression of NF-kB and its down-stream antiapoptosis gene in HCC, Bcl-2, with occurrence and development, the relation each other. Observe the change of NF-kB in cytoplasm and nucleus induced by chemotherapy drug, doxorubicin. Investigatite the role that NF-kB and its specific inhibitor plays in the liver cancer cells apoptosis , explore the reason of liver cancer cells persisting in apoptosis induced by chemotherapy drug and look for thecorresponding methods to inhibit so as to offer the test basis to improvetreatment efficiency of chemotherapeutic drug.AIM:To explore NF-kB expression in liver carcinoma tissues and its correlation with the pathological stage of cancer tissues and the expression of Bcl-2.To explore that NF-kB transfers from cytoplasm to nucleus of hepatocellular carcinoma (SMMC-7721) after activated by doxorubicin. After NF-kB was inhibited by the specific inhibitor of NF-kB, PDTC, the apoptosis index of SMMC-7721 cells changed during the period. METHODS:By means of immunohistochemical SABC method, the expressions of NF-kB and Bcl-2 were compared in hepatocellular carcinoma tissues from 40 cases and normal liver tissues from 15 cases. The transferring of NF-kB from cytoplasm to nucleus was observed by means of immunohistochemical SABC method. After the nucleus protein of SMMC-7721 cells had been extracted, ELISA assay was performed to detect the change of NF-kB in the nucleus treated by doxorubicin or doxorubicin plus PDTC. The death rate and the apoptosis index of SMMC-7721 cells on different conditions were detected by MTT assay and Tunel method individually. RESULTS:The positive expression of NF-kB in the liver cancer group and the control group were 72.5%,13% (signinificantly different P<0.01) and intensive expression of NF-kB were 30%, 7% respectively. The positive expression of Bcl-2 were 40%,7% (signinificantly different P<0.05)and the intense expression of Bcl-2 were 17.5%, 0% respectively. The positive expression of NF-kB was significantlydifferent for different pathological grades (P<0.01). The expression of Bcl-2 was closely related to NF-kB (r=0.389, P<0.01). The transferring from cytoplasm to nucleus of NF-kB induced by doxorubicin increased gradually with time. After the nucleus protein of SMMC-7721 cells had been extracted, ELISA assay was performed to detect the change of NF-kB in the nucleus treated by doxorubicin or doxorubicin plus PDTC in 3h,6h, 12h. OD was 0.106+0.25 0.29+0.03 0.83 +0.14 and 0.005 +0 0.0095+0 0.014 + 0 individually. MTTassay: IC50(doxorubicin) =0.16, IC50(doxorubicin plus PDTC) =0.06. 24h 48h 72h apoptosis index of doxorubicin group and doxorubicin plus PDTC group were 0.025 0.05 0.15 and 0.16, 0.43 0.49 individually. CONCLUSION:The abnormal expression of NF-kB is closely associated with the development of liver cancer. The expression of NF-kB is significantly different for different pathological grades. Its high expression indicates a poor promising. NF-kB is closely related to Bcl-2, which indicates that NF-kB is involved in the anti-apoptosis course of liver cancer cell by mediating anti-apoptosis gene.NF-kB in SMMC-7721 cells can be activated by DNA-damage drug such as doxorubicin and transfers from cytoplasm into nucleus, which can be specially i... |
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