Transfection of antisense c-myc into human glioma U251 cells by dint of non-phospholipid liposome
Objective To explore the effect of antisense c-myc on human glioma cell U251 after transfection by dint of non-phospholipid liposome (NPL). Methods pHMTASCMH, which inducibly expresses human antisense c-myc under the control of human metallothionein promoter, was mixed with NPL and added into U251 cell culture. ZnCl 2 (200μmol/L) was added after 6 hours. 3-[4,5-dimethylthiazolzyl]-2,5-diphenyl tetrazolium bromide (MTT) test was used to evaluate cell activity after 24-96 hours of the antisense c-myc transfection. The cells treated for 24 or 72 hours were microscoped in fluoresce microscope after Annexin-V-FLUOS staining or TUNEL staining respectively. The treated cells were lysed and then c-myc protein expression was assayed by ELISA. Results Complex of antisense c-myc and NPL significantly inhibited the proliferation of glioma cells in the presence of Zn 2+. NPL did not significantly affect cell growth at the concentration tested. The treated cells lost normal morphology after 24 hours and cytoplasmic vacuoles were found. Phenomena correlative with programmed cell death were well-identified in treated cells after Annexin-V-FLUOS or TUNEL staining. ELISA demonstrated the decrease of c-myc expression products. Conclusion Antisense c-myc inhibited the expression of c-myc, and proliferation of glioma cells and induced cell death.