The Effects Of Soluble CD40Ligand On THP-1Cell Proliferation And The Expression Of Bcl-2, P53mRNA

Objective:To observe the effect of sCD40Lon the cell proliferation, cell death, expressionof Bcl-2and P53of acute monocytic leukemia (M5）cell line THP-1cells．Methods:（1）THP-1cell were co-incubated with sCD40L of different concentrations.CCK-8assay was used to detect the cell proliferation and chose the best concentration ofsCD40L.(2) Flow cytometry was used to examine their apoptosis.(3) RT-PCR was used todetermine Bcl-2and P53mRNA expression.Results:(1)Compare with THP-1cell groups, sCD40L were able to inhibit THP-1cellsproliferation in a dose-dependent manner for24、48、72hours(P<0.05), The increase ofgrowth inhibitory rate was most significant at its concentration of4.00μg/ml. sCD40Lwere co-incubated with on THP-1cells for24h,48h,72h,all had different degree ofinhibition proliferation in vitro and sCD40L4.00μg/ml was the strongest at the sametime(P<0.05).(2)sCD40L effected on THP-1cells for three different time can be asignificant apoptosis,compared with cell group(P<0.05), The apoptosis rate was mostsignificant for72h.(3) THP-1cells cultured with sCD40L, In THP-1cells cultured withsCD40L4.00μg/ml for the same time the expression of Bcl-2mRNA was down-regulatedmost obviously and P53expression was up-regulated most significantly at the sametime(P<0.05), two genes were related to time at the different time.Conclusion:(1) sCD40L can induce growth inhibition and apoptosis in THP-1cells, itwas related to concentration and time;(2) In the process of THP-1cells apoptosis, themitochondria pathway plays an important role. In another words,the molecular mechanismof THP-1cells apoptosis induced by sCD40L is related to the down-regulation of Bcl-2and up-regulation of P53.