关键词: 端粒长度, 端锚酶, 端粒酶催化亚单位, 反义寡核苷酸, 人肺腺癌A549细胞

Abstract: Objective The aim of this study is to determine the effect of transcription and translation in telomeric related proteins, and synergism of progressive telomere shortening and cell cycle alteration in human lung adenocarcinoma A549 cell lines, which induced by antisense tankyrase oligonucleotide(asTANKS) combinated with antisense human telomerase reverse transcriptase (ashTERT) oligonucleotide. Methods A549 cells was randomly assigned to 3 groups: ashTERT, ashTERT + asTANKS and asTANKS, while 3 groups (shTERT, sTANKS and blank) as control. With individual intervention for different hours, the effect of transcription in hTERT mRNA was evaluated by RT-PCR, and telomerase activity by ELISA-PCR, tankyrase activity by Western Bloting as well. Moreover, telomere average length was analyzed by Q-FISH, and besides, duration of proliferation was observed by population double test. Results Transcription in hTERT mRNA and telomerase activity for 48h was inhibited obviously by ashTERT , but little done by asTANKS which abated significantly tankyrase activity. Progressive telomere shortening in A549 cells for 48h was evident induced by either asTANKS or ashTERT (Vs control, P<0.01), but it was more severe by combination of them (Vs ashTERT / asTANKS, P<0.01). Furthermore, continuous treatment of ashTERT was similar to asTANKS in duration of proliferation, which was observed 53-57PD and 56-58PD respectively (Vs control, P<0.01), and combinative effect of them conduced to a shorter survival (22-26PD) and earlier cell crisis onset (Vs ashTERT / asTANKS, P<0.01). Conclusions As has been mentioned above, distinguished from other inhibitor concerned with dynamics of telomerase, asTANKS does not only lead to progressive telomere shortening, but also work in coordination with ashTERT in A549 cell lines, which enhanced the efficacy of telomere shortening and hastened earlier cellular crisis. This study provides insight into strategies for telomere-based molecular cancer therapeutics.

Key words: telomere length, tenkyrase (TANKS), human telomerase reverse transcriptase (hTERT), antisense oligonucleotide (ASODN), human lung adenocarcinoma A549 cell lines