ABSTRACT
Objective@#To further understand the interaction protein spectrum of heterogeneous ribonucleoprotein AB (hnRNP AB), and to investigate their clinical significance in hepatocellular carcinoma (HCC).@*Methods@#We carried out mass spectrometry to reveal the specific peptides of KRAB-associated protein 1 (Kap1) and hnRNPAB, and verified their interaction by immunocoprecipitation and western blotting. Expression of hnRNPAB/Kap1 proteins were detected by immunohistochemical staining in the tissue microarrays. Categorical data were analyzed by the chi square test or Fisher exact test; enumeration data between groups were compared using Student t-test or Wilcocon signed rank test; the cumulative recurrence and survival rates were evaluated using the Kaplan-Meier method and the differences were assessed using the log-rank test.@*Results@#We identified Kap1 as a molecular partner for hnRNPAB in HCCLM3 cells and HepG2 cells as well. We found that the 5-year survival rate of the Kap1high patients was significantly lower than the survival rate of those of the Kap1low group (36% vs 59% , HR = 1.67, P < 0.001). Similarly, Kap1high HCC patients had the poorest prognosis at 5-years, with higher cumulative recurrence rate than Kap1low patients (72% vs 54%, HR = 1.66, P = 0.001). Univariate and Multivariate analyses revealed that hnRNPAB /Kap1 alone (HR = 1.35 /1.28, P = 0.001) or in combination with Kap1 (HR =1.24 /1.27, P < 0.05) were independent prognostic indicators for overall survival and time to recurrence.@*Conclusion@#In HCC cells, hnRNPAB and Kap1 form protein complexes. The expression levels of hnRNPAB alone or in combination with Kap1 in HCC patients are important because they provide not only a predictor for HCC prognosis but also a therapeutic target for future studies.
ABSTRACT
Objective:To observe the effects of isobavachalcone (IBC)on the proliferation and apoptosis of tongue squamous cell carcinoma Tca-8113 cells,and to explore their mechanisms. Methods:The Tca8113 cells cultured in vitro were divided into control group and different doses (0, 10, 20,40, 80 μmol · L-1 )of IBC groups.The inhibitory rates of cell proliferation were detected by MTT method.The apoptosis was detected by flow cytometry.Western blotting method was used to detect the expressions of Akt,p-Akt,Erk,p-Erk,Bax, Bcl-2 and Caspase-3 proteins in the Tca8113 cells in various groups.Results:The MTT results showed that the inhibitory rates of proliferation of Tca8113 cells were increased in a concentration-and time-dependent manner;the IC50 at 12,24 and 48 h were (285.13±8.97), (132.40±7.76),and (58.56±5.93)μmol·L-1 ,respectively;and there were significant differences between different time points (P 0.05 ).The expression levels of Caspase-3 in Tca8113 cells in 40 μmol·L-1 IBC group at 48 h were increased compared with control group (P <0.05).Conclusion:IBC could inhibit the proliferation and induce the apoptosis of Tca8113 cells;Akt and Erk signaling pathway may be the pathway of IBC to induce the apoptosis of tumor cells.
ABSTRACT
Aim To explore the inhibitory effect of isobavachalcone (IBC)on migration and invasion of tongue squamous cell carcinoma Tca81 1 3 cells and its possible mechanism.Methods Tca81 1 3 cells were treated in different concentrations of IBC in vitro.Cell proliferation was detected by MTT;Wound healing as-say and Transwell chamber assay were used to detect the ability of cell migration and invasion;Western blot was applied to detect the expression of Akt,p-Akt, MMP-2 and MMP-9 proteins.Results IBC could in-hibit the proliferation of Tca81 1 3 cells in a concentra-tion-and time-dependent manner.IBC can reduce cell migration and invasion.Western blot showed that IBC could an decrease the expression of p-Akt,MMP-2 and MMP-9 proteins in a concentration-dependent manner. However,the level of Akt was not affected by the con-centration of IBC treatment.Conclusion IBC could inhibit the proliferation, migration and invasion in Tca81 1 3 cells and its mechanism may be associated with the down-regulation of MMP-2 and MMP-9 pro-teins and the inhibition of phosphorylation of upstream Akt.
ABSTRACT
<p><b>OBJECTIVE</b>To investigate the effect of 17-allylamino-demethoxygeldanamycin (17AAG) on the proliferative and invasive ability of gastric cancer cells and associated mechanism.</p><p><b>METHODS</b>The proliferative ability was tested by MTT method and the cell cycle was detected by flow cytometry(FCM) when 17AAG was used to treat gastric cancer cell SGC7901. Apoptosis was detected by FCM and PI-Annexin V double staining. The invasive ability was tested by transwell method. Expression of HSP90, HSP70, c-met and AKT was detected by Western blot.</p><p><b>RESULTS</b>The growth of SGC7901 cells was inhibited after the administration of 17AAG, and the inhibitation was dose- and time-dependent. The cell cycle was blocked at the G0/G1 phase. The apoptotic ratio in 17AAG group was much higher than that in blank group and DMSO group (P<0.01). The cellular invasive ability decreased significantly (P<0.01). The expression of HSP70 was elevated by 17AAG, and the expression of c-met and AKT was down-regulated, but no change of HSP90 was observed.</p><p><b>CONCLUSION</b>17AAG can inhibit the proliferative and invasive ability of SGC7901 cells, and induces apoptosis through down-regulating the expression of HSP90 client proteins instead of the target HSP90 itself.</p>
Subject(s)
Humans , Apoptosis , Benzoquinones , Pharmacology , Cell Cycle , Cell Line, Tumor , Cell Proliferation , HSP70 Heat-Shock Proteins , HSP90 Heat-Shock Proteins , Lactams, Macrocyclic , Pharmacology , Neoplasm Invasiveness , Stomach Neoplasms , PathologyABSTRACT
Objective To observe the expressions ofα-catulin and E-cadherin in tongue squamous cell carcinoma (TSCC)tissue, and to explore their relationship with the occurrence and development of TSCC.Methods The expressions ofα-catulin and E-cadherin in 55 cases of TSCC tissue and 10 cases of normal tongue tissue were examined by immunohistochemistry SP method. The relationship between the expressions and the clinicopathological characteristics and the relevance of the expressions ofα-catulin and E-cadherin in TSCC tissue were analyzed.Results The positive expression rates ofα-catulin in TSCC tissue and normal tongue tissue were 69.09% and 20.00%,respectively,and there was difference between them (P<0.01).The expression ofα-catulin was correlated to the histological differentiation,clinical stage and lymph node metastases of TSCC (P<0.05).The positive expression rates of E-cadherin in TSCC tissue and normal tongue tissue were 38.18% and 80.00%,respectively, and there was significant difference between them (P< 0.01 ). The expression of E-cadherin was correlated to the clinical stage and lymph node metastases of TSCC (P<0.05 ). There was a negative correlation between the expressions ofα-catulin and E-cadherin in TSCC tissue (r=-0.466,P<0.01). Conclusion The expressions ofα-catulin and E-cadherin may be associated with the occurrence and development of TSCC,and they could be used as the parameters which predict the malignant degree and prognosis of TSCC.
ABSTRACT
Highly active adherent LAK cells (A-LAK) with monocytes depleted by phenylalahine methyl ester (PME) were cultured from peripheral blood lymphocytes of patients with hepatocellular carcinoma (HCC). Results showed that A-LAK cells cultured in about 14 days had expanded better and faster than that of nonadherent LAK cells (NA-LAK) with their greatest expansion varied from 23 to 243 fold .A-LAK cells showed a trend of increase in TH cell subgroup and decrease in Ts cell sub-group as well as significant difference of TH/TS ratio. The IL-2R expression increased from 34.1%to 64.3%. A-LAK cells had a higher cytotoxicity (64.6%)than that of NA-LAK cells (42.8%).Further clinical application of A-LAK cells may improve biotherapeutic effect on HCC patients compared with that of NA-LAK cells .
ABSTRACT
To identify the best inducing condition, we studied the expression of membrane protein HSP70 and mRNA of H22 cell at various temperature. Using MTT,RT-PCR, immunofluorescence and FCM techniques, we observed H22 cell survival rate, the expression of HSP70 mRNA and membrane HSP70. No effects of H22 cell survival rate under 42 ~ 43℃ was observed, but cell survival rate declined with increasing stress time at 44 ~ 45 ℃; the level of HSP70 mRNA decreased initially (0.5~4.0) hours but gradually resumed and increased from 8 to 12 hours at 42℃. Membrane HSP70 expressing cells were significently higher in heat shock treatment group than in a control group ( P