ABSTRACT
Objective To investigate the expression of c-FLIPL in leukemia and explore its clinical significance. Methods The expression level of c-FLIPL mRNA in bone marrow mononuclear cells was determined by real-time polymerase chain reaction(PCR)in 103 leukemia patients with different types of leukemia,including 54 cases of acute lymphocytic leukemia(ALL)with 37 newly diagnosed,5 relapsed,and 12 complete remis-sion,38 cases of acute myeloid leukemia(AML)with 24 newly diagnosed,6 relapsed,and 8 complete remission,newly diagnosed 2 cases of acute undifferentiated leukemia(AUL),6 cases of chronic myelocytic leukemia(CML),and 3 cases of chronic myelomonocytic leukemia(CM-ML). The immunophenotype of patients were detected by flow cytometry. Results Expression level of c-FLIPL mRNA was higher in newly diag-nosed and relapsed leukemia patients. There was no significant difference between newly diagnosed and relapsed leukemia patients(P>0.05). Ex-pression level of c-FLIPL mRNA of AUL and CML was higher than that in other patients ,but there was no significant difference(P>0.05). Ex-pression level of c-FLIPL mRNA of all newly diagnosed and relapsed leukemia patients was significantly higher than that in control group and com-plete remission group(P<0.05). The expression level of c-FLIPL mRNA was correlated with risk stratification ,white blood cell(WBC),lactate dehydrogenase(LDH),hydroxybutyrate dehydrogenase(HBDH),CD45 and TEL-AML1,but was not associated with age,sex,fibrinogen and chromosome abnormality. Conclusion c-FLIPL mRNA is highly expressed in leukemia patients ,and is closely related with risk stratification , WBC,LDH,HBDH and prognosis.
ABSTRACT
Overexpression of Short and Raji variants of Cellular FLICE-like inhibitory protein (c-FLIP) is capable of inhibiting apoptosis, while the function of the Long isoform depends of c-FLIPL concentration in cells. The aim of this study was to determine the effects of c-FLIPL knockdown in cervical cell lines. SiHa, C-4I and C-33A cervical cancer cell lines were analyzed. c-FLIPL level expression was determined by quantitative real-time PCR and western blotting. c-FLIPL was transiently downregulated by siRNA. The effects of knockdown of c-FLIPL on cell viability, proliferation and apoptosis were assessed by comparing with scrambled siRNA-transfected cells. SiHa and C-4I c-FLIPL knockdown cells showed increased viability compared with scrambled siRNA-transfected cells (P<0.05), while C-33A cells did not show significant differences. Ki-67 and PCNA immunocytochemistry was performed to evaluate proliferation on these cervical cancer cell lines. SiHa cells with c-FLIPL knockdown showed elevated expression of Ki-67 protein compared with their scrambled counterparts (P<0.0001), while C-33A c-FLIPL knockdown cells showed a significantly lower in PCNA expression (P<0.01) compared with control. All three c-FLIP-transfected cell lines showed a higher level of apoptosis compared with their scrambled controls. Our results suggest that c-FLIPL could have effects in proliferation and apoptosis in cervical cancer cell lines.
Cuando las variantes Short y Raji de la proteína Cellular FLICE-like inhibitory protein (c-FLIP) se encuentran sobrexpresadas son capaces de inhibir la apoptosis, mientras la función de la isoforma Long (c-FLIPL), depende de la concentración de esta molécula en las células. El objetivo de este estudio fue determinar los efectos de la inhibición de c-FLIPL en líneas celulares de cáncer de cuello uterino. Para realizar el estudio fueron utilizadas SiHa, C-4I y C-33A, líneas celulares de cáncer cervical. La expresión de c-FLIPL en estas líneas fue establecida mediante PCR en tiempo real y western blot. Posteriormente la expresión de c-FLIPL fue inhibida, mediante transfeción transiente con siRNA complementario al mRNA mensajero de c.-FLIPL. Los efectos de esta inhibición en la viabilidad celular, proliferación y apoptosis fue comparada con células transfectadas con un siRNA control (scrambled). Una vez reprimido c-FLIPL, las líneas celulares SiHa y C-4I presentaron un aumento de la viabilidad celular (P<0,05). Para evaluar la proliferación celular se utilizó inmunocitoquímica de los marcadores Ki-67 y PCNA. Las células SiHa transfectadas con siRNA c-FLIPL, mostraron una elevada expresión de Ki-67 (P<0,0001), mientras que las células C-33A con c-FLIPL inhibido mostraron una menor expresión de PCNA (P<0,01). Las tres líneas celulares con c-FLIPL reprimido mostraron un mayor nivel de apoptosis que las células control. Estos resultados sugieren que c-FLIPL puede tener efectos en la proliferación y apoptosis de líneas celulares de cáncer de cuello uterino.