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Expression and biological significance of c-FLIP in human hepatocellular carcinomas.

Du X, Bao G, He X, Zhao H, Yu F, Qiao Q, Lu J, Ma Q - J. Exp. Clin. Cancer Res. (2009)

Bottom Line: And doxorubicin showed apparent inhibition on cell proliferations, and induced more apoptosis.These results indicate that c-FLIP is frequently expressed in human HCCs, and its overexpression implied a lesser probability of recurrence-free survival.The specific silencing of c-FLIP gene can apparently up-regulate drug-induced HCC cell apoptosis, and may have therapeutic potential for the treatment of human HCC.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of General Surgery, Tangdu Hospital, Fourth Military Medical University, Xi'an 710038, PR China. tdsurg@fmmu.edu.cn

ABSTRACT

Background: c-FLIP can be considered as a tumor-progression factor in regard to its anti-apoptotic functions. In the present study, we intended to investigate the expression of c-FLIP in human HCC tissues, and its relation with drug-induced cell apoptosis through the specific inhibition of c-FLIP expression by siRNA in 7721 cells.

Methods: c-FLIP expression was quantified immunohistochemically in HCC tissues(eighty-six cases), and corresponding noncancerous tissues (fifty-seven cases). Patients with HCC were followed up for cancer recurrence. Then, the c-FLIP gene was silenced with specific siRNA in 7721 HCC cells. c-FLIP expression was detected by RT-PCR, Western Blot and immunocytochemical staining. The cellular viability and cell apoptosis were assayed in vitro with cells treated with doxorubicin.

Results: Positive immunostaining was detected for c-FLIP in 83.72% (72/86) human HCC tissues, 14.81% (4/27) hepatic cirrhosis, 11.11% (2/18) hepatic hemangioma tissues, and absent in normal hepatic tissues. The overexpression(more than 50%) of c-FLIP in HCC adversely affected the recurrence-free survival. Through c-FLIP gene silencing with siRNA, the expressions of c-FLIP mRNA and protein were remarkably down-regulated in 7721 HCC cells. And doxorubicin showed apparent inhibition on cell proliferations, and induced more apoptosis.

Conclusion: These results indicate that c-FLIP is frequently expressed in human HCCs, and its overexpression implied a lesser probability of recurrence-free survival. The specific silencing of c-FLIP gene can apparently up-regulate drug-induced HCC cell apoptosis, and may have therapeutic potential for the treatment of human HCC.

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Cell viability was accessed by cell counting. The study showed that 7721 cell viability was reduced by the transfetion with recombinant iRNA vectors. pSuper-Si1 had more significant effect on the reduction of the cell viability.
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Figure 4: Cell viability was accessed by cell counting. The study showed that 7721 cell viability was reduced by the transfetion with recombinant iRNA vectors. pSuper-Si1 had more significant effect on the reduction of the cell viability.

Mentions: Cells were treated with doxorubicin for 0–8 days, and then the growth curves were obtained. Cell proliferation was inhibited obviously when c-FLIP expression was knocked down by siRNA. Our data showed that si-526-siRNA significantly decreased the growth rate of 7721 cells, with a >50% decrease after 3 days repeatedly in three separate experiments (Figure. 4).


Expression and biological significance of c-FLIP in human hepatocellular carcinomas.

Du X, Bao G, He X, Zhao H, Yu F, Qiao Q, Lu J, Ma Q - J. Exp. Clin. Cancer Res. (2009)

Cell viability was accessed by cell counting. The study showed that 7721 cell viability was reduced by the transfetion with recombinant iRNA vectors. pSuper-Si1 had more significant effect on the reduction of the cell viability.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2654864&req=5

Figure 4: Cell viability was accessed by cell counting. The study showed that 7721 cell viability was reduced by the transfetion with recombinant iRNA vectors. pSuper-Si1 had more significant effect on the reduction of the cell viability.
Mentions: Cells were treated with doxorubicin for 0–8 days, and then the growth curves were obtained. Cell proliferation was inhibited obviously when c-FLIP expression was knocked down by siRNA. Our data showed that si-526-siRNA significantly decreased the growth rate of 7721 cells, with a >50% decrease after 3 days repeatedly in three separate experiments (Figure. 4).

Bottom Line: And doxorubicin showed apparent inhibition on cell proliferations, and induced more apoptosis.These results indicate that c-FLIP is frequently expressed in human HCCs, and its overexpression implied a lesser probability of recurrence-free survival.The specific silencing of c-FLIP gene can apparently up-regulate drug-induced HCC cell apoptosis, and may have therapeutic potential for the treatment of human HCC.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of General Surgery, Tangdu Hospital, Fourth Military Medical University, Xi'an 710038, PR China. tdsurg@fmmu.edu.cn

ABSTRACT

Background: c-FLIP can be considered as a tumor-progression factor in regard to its anti-apoptotic functions. In the present study, we intended to investigate the expression of c-FLIP in human HCC tissues, and its relation with drug-induced cell apoptosis through the specific inhibition of c-FLIP expression by siRNA in 7721 cells.

Methods: c-FLIP expression was quantified immunohistochemically in HCC tissues(eighty-six cases), and corresponding noncancerous tissues (fifty-seven cases). Patients with HCC were followed up for cancer recurrence. Then, the c-FLIP gene was silenced with specific siRNA in 7721 HCC cells. c-FLIP expression was detected by RT-PCR, Western Blot and immunocytochemical staining. The cellular viability and cell apoptosis were assayed in vitro with cells treated with doxorubicin.

Results: Positive immunostaining was detected for c-FLIP in 83.72% (72/86) human HCC tissues, 14.81% (4/27) hepatic cirrhosis, 11.11% (2/18) hepatic hemangioma tissues, and absent in normal hepatic tissues. The overexpression(more than 50%) of c-FLIP in HCC adversely affected the recurrence-free survival. Through c-FLIP gene silencing with siRNA, the expressions of c-FLIP mRNA and protein were remarkably down-regulated in 7721 HCC cells. And doxorubicin showed apparent inhibition on cell proliferations, and induced more apoptosis.

Conclusion: These results indicate that c-FLIP is frequently expressed in human HCCs, and its overexpression implied a lesser probability of recurrence-free survival. The specific silencing of c-FLIP gene can apparently up-regulate drug-induced HCC cell apoptosis, and may have therapeutic potential for the treatment of human HCC.

Show MeSH
Related in: MedlinePlus