Research on anti-sense nucleotide technology in current lung cancer treatment / 肿瘤研究与临床

Lung cancer has been widely known for its highest morbidity among all tumors in this world.Anti-sense neoclitide technology is a kind of widely-recognized gene therapy for its high efficiency and practical simplicity.This essay will briefly review principles of anti-sense neoclitide technology used in treatment of lung cancer and characteristics of this technology in gene therapy of lung cancer.

Effect of antisense oligodeoxynucleotide on cell proliferation and telomerase activity in K562 cell line / 白血病·淋巴瘤

Objective To observe anti-sense phosphorothioate oligonucletide (ASPSODN) targeted directly to hTERT mRNA to its inhibiting effect on aimed gene and the influence on the telomerase activity, cellular proliferation, cell apoptosis of K562 cells. Methods Human leukemia cell line K562 was transfected with anti-sense oligonucleotide ASPSODN by liposome. The proliferation activity of K562 cell line was determined by using methyl thiazolyl tetrazolium assay, and telomerase activity was detected by TRAP-PCR-ELISA. Flow cytometry was adopted to examine apoptotic rate and cell cycle. RT-PCR was used to detect the expression of target gene hTERT mRNA. Results 0.6 μmol/L ASPSODN (0.42 ±0.16) was remarkably decreased the expression of hTERT mRNA, Telomerase relative activation was decreased by 52 %. According to 0.6 (μmol/L ASPSODN caused significant the inhibition of K562 cell growth. Apoptotic rate and cell cycle was examined by 0.6 μmol/L ASPSODN with flow cytometry. The cell apoptosis rate of 0.6μmol/L ASPSODN were 10.31 %. It showed the cells treated with 0.6 uU PSASODN arrested in G_1/C_0. The ratio of cells in G_2/M and S period was reduced. But there was no characteristic apoptosis peak. Conclusion ASPSODN targeted hTERT can inhibit the expression of target gene hTERT mRNA, and decrease the telomerase activity of K562 cells. ASPSODN can inhibit strongly the proliferation of K562 cell and induce cell apoptosis by decreasing telomerase activity.

Influence of MMP-9 antiseme oligonudeotide on proliferation and apoptosis of human lung adenocarcinom A549 cells / 中国医师杂志

Objective To approach the effects of MMP-9 antisensc oligonucleotide(ASODN)on human lung adcnocarcinoma(A549) cell apoptosis and proliferation capability.Methods MTT.was used to analyze the effect that MMP-9 transfection on A549 cell growth.Flow cytometry was used to analyze the ratio of cell proliferation and apoptosis.RT-PCR WaS used to detect the expression of MMP-9mRNA and Western blot was used to detect the changes of MMP-9 protein.Results In some degree,MMP-9 ASODN that inhibited the survival rate of the A549 cell presented concentration and time dependence manner.The best dependence concentration of ASODN was 600nmol/L for 24 hour.After trasnfection of MMP-9ASODN to A549 cell.the perceive of A549 apoptosis cell was significandy higher than that in control group(P＜0.01).When the antisense oligonucleotides concentration is 600nmol/L and the action time is 48 hours,the relative expression level of MMP-9mRNA and MMP-9 protein are obviously less than that in control group(P＜0.01).Conclusion MMP-9ASODN may down regulate the expression of MMP-9mRNA and MMP-9 protein,effectively inhibit the proliferation of A549 cell and promote the apoptosis of A549 cell.

Antisense oligonucleotide mediated inhibition on telomerase activity in gallbladder carcinoma cell / 中国现代普通外科进展

Objective:To study the antisense oligonucleotide mediated inhibition on telomerase activity and cell proliferation of GBC-SD cell.Methods:We design the antisense,sense,and random oligonucleotide with phosphoric acid modification for the hTR(Human Telomerase RNA)template sequence.MTT and PCR methods were used to observe the inhibition on telomerase activity and cell proliferation of GBC-SD cell ,and fibroblast cells were used as control group.Results:PS-ODN can lead to the reduction of cell survival rate of GBC-SD cell,wich dosage dependence.Tne experimental group cell detected by scanning electron appeared apoptotic feature.Conclusion:PS-ODN can inhibit telomerase activity of GBC-SD cell effectively and induce the cell apoptosis.

Effect of ultrasound-mediated triplex-forming oligonucleotide lipid ultrasonic microbubble on cellular absorption rate of TFO and expression of tissue factor / 解放军医学杂志

Objective To investigate the effect of lipid ultrasonic microbubble on the cellular absorption rate of antiparallel phosphorothioate triplex-forming oligonucleotide(TFO) and the expression of the tissue factor(TF) in endothelial cells.Methods The GT21-apsTFO,specific to the gene sequence of the shearing stress response element,was synthesized and the lipid ultrasonic microbubble carrying TFO was constructed.The ECV304 cells were divided into 4 groups:control group(SSRE),received shear stress only for 6h;TFO+ ultrasonic irradiation group(TFO+U),treated with 60?l TFO in final concentration of 0.2?mol/L and ultrasonic irradiation for 30s simultaneously;TFO-lipid microbubble group(TFO-M),treated with 60?l lipid microbubble loaded with TFO in final concentration of 0.2?mol/L;and TFO-lipid microbubble + ultrasonic irradiation group(TFO-M+U),treated with 60?l lipid microbubble loaded with TFO(final concentration was 0.2?mol/L) and ultrasonic irradiation for 30s simultaneously.Cells in the last 3 groups were treated with shear stress for 6h with pressure of 1.2Pa 24h after transfection.The fluorescent microscope was used to observe the cellular absorption rate of TFO.The immunofluorescence method and RT-PCR were employed to determine the protein and mRNA expressions of TF.Results The transfection efficiency of the TFO was higher in TFO-M+U group(38.83%?6.52%) than in TFO-M group(9.50%?2.88%) and TFO+U group(12.66%?3.01%,P

Effects of antisense vascular endothelial growth factor-C gene transinfection on tumorigenicity and vascular generation of human gastric carcinoma cells / 解放军医学杂志

Objective To investigate the biological effects of human gastric carcinoma cell line SGC-7901 transinfected by antisence vascular endothelial growth factor-C (anti VEGF-C) gene on the tumorigenicity and vascular generation,and explore the role of VEGF-C gene in metastasis and vascular generation of gastric cancer.MethodsThirty nude mice were randomly divided into three groups (10 each):transgenic anti VEGF-C group,transgenic pure liposome group and non-transgenic group.0.2ml (1?107/ml) of SGC-7901 suspension,either transinfected by anti VEGF-C gene,or by pure liposome,or transinfected nil,was respectively subcutaneously injected into the three groups of mice,once every 2 days for 3 consecutive days.After injection,the size and growth velocity of the neoplasm were measured.The microlymphatic vessel density (MLVD) and microvessel density (MVD) of tumor were also determined.ResultsThe neoplasm grew more slowly and was smaller in the mice of transgenic anti VEGF-C group than in non-transgenic ones.At the end of the 1st,2nd and 3rd week,the MLVD in the mice of transgenic anti VEGF-C group was 4.0?2.2,6.0?3.1 and 9.0?2.7/high field (/HF),respectively.In the transgenic pure liposome group,the MLVD was 6.0?8.7,9.0?3.5 and 18.0?7.2/HF,respectively.In the non-transgenic group,the MLVD was 7.0?4.9,9.0?6.4 and 19.0?6.5/HF,respectively.The MVD in the transgenic anti VEGF-C group was 3.0?2.4,5.0?2.1 and 8.0?1.7/HF,respectively.In the transgenic pure liposome group was 4.0?1.8,6.0?2.7 and 10.0?1.3/HF,respectively,and in non-transgenic group was 4.0?1.5,6.0?1.3 and 9.0?1.2/HF,respectively.There existed significant difference in lymphangiogenesis of the tumor (P0.05).ConclusionsThe anti VEGF-C gene may suppress the tumorigenicity and lymphangiogenesis of human SGC-7901 gastric carcinoma cells,but have little effect on angiogenesis of the tumor.VEGF-C might participate in the lymphangiogenesis of gastric tumor.