Wang Zhiping,Zhou Jianchao,Zhang Zhenzhong,Fu Lei.PEI/ASON-mediated nanoparticle assembly suppresses proliferation and hTERT mRNA expression in lung cancer cell line A549[J].J Amry Med Univ (J Third Mil Med Univ),2012,34(08):749-753.
Wang Zhiping,Zhou Jianchao,Zhang Zhenzhong,Fu Lei.PEI/ASON-mediated nanoparticle assembly suppresses proliferation and hTERT mRNA expression in lung cancer cell line A549[J].J Amry Med Univ (J Third Mil Med Univ),2012,34(08):749-753.DOI:
PEI/ASON-mediated nanoparticle assembly suppresses proliferation and hTERT mRNA expression in lung cancer cell line A549
Objective To determine the effect of nanoparticles for antisense oligodeoxynucleotide (ASODN) of human telomerase reverse transcriptase (hTERT) mRNA on the proliferation and hTERT expression in A549 cells. Methods MTT assay was used to detect the proliferation in A549 cells
and the cells treated by ASODN
PEI/SODN and PEI/ASODN (40 μg/ml). Intracellular fluorescence intensity was detected by laser confocal microscopy in the cells after being transfected with 5′-FITC-labelled ASODN (FASODN). Inverse microscopy was used to observe the modality of A549 cells transfected by PEI/ASODN. The expression of hTERT mRNA were determined by RT-PCR. The expression of hTERT protein was detected by immunohistochemictry assay kit. Cell apoptosis were determined by flow cytometry (FCM). To observe the tumor targeting effect of NGR (Asn-Gly-Arg)
a total of 36 A549 cells transplanted nude rats were injected with ASON
PEI/ASON
PEI/ASON-PEG and PEI/ASON-PEG-CNGR solutions respectively via tail vein. The distribution of PEI/ASON-PEG-CNGR in the heart
brain
kidneys
lungs and liver was observed in 1
3 and 6 h after injection. Results PEI/ASODN exerted inhibitory effects on the proliferation in A549 cells in a time-dependent manner. The intracellular fluorescence in PEI/FASODN treated cells was obviously stronger than that in FASODN group (PEI free) after transfection. After transfecting PEI/ASODN for 72 h
the hTERT mRNA expression was decreased obviously compared with control group (P0.05). The hTERT protein expression of PEI/ASODN group was reduced. FCM showed that the early and later apoptosis rates in PEI/ASODN group was 36.53% and 54.28% respectively. Animal in vivo experiment showed that PEI/ASON-PEG-CNGR had good tumor targeting effect. Conclusion PEI/FASODN of hTERT can inhibit the proliferation in A549 cells effectively
suppress mRNA and protein expression of hTERT and cause cell apoptosis.