Effects of Recombinant Adenovirus Carrying Apoptin Gene on the Apoptosis of Human Lung Squamous Carcinoma SK-MES-1 Cell Lines and Human Lung Adenocarcinoma NCI-H1299 Cell Lines
10.6039/j.issn.1001-0408.2015.22.13
- VernacularTitle:携带凋亡素基因的重组腺病毒对人肺鳞癌细胞SK-MES-1、人肺腺癌细胞NCI-H1299凋亡的影响
- Author:
Jun RUAN
;
Youru WU
;
Lili MIAO
- Publication Type:Journal Article
- Keywords:
Apoptin gene VP3;
Recombinant adenovirus vector Adxsi-GFP-VP3;
Human lung squamous carcinoma SK-MES-1 cell lines;
Human lung adenocarcinoma NCI-H1299 cell lines;
Protein expression;
Apoptosis;
Cell cycle
- From:
China Pharmacy
2015;(22):3066-3069
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE:To explore the effects of recombinant adenovirus vector Adxsi-GFP-VP3 carrying apoptin gene VP3 on the apoptosis of human lung squamous carcinoma SK-MES-1 cell lines and human lung adenocarcinoma NCI-H1299 cell lines. METHODS:The exponential phase SK-MES-1 and NCI-H1299 cell lines were respectively divided into a recombinant adenovirus (Adxsi-GFP-VP3) group,a empty virus (Adxsi-GFP) group and a cell control (phosphate buffer) group,which were marked as group A,B and C respectively. Reverse transcription polymerase chain reaction and Western blot method were used to detect the ex-pressions of VP3 mRNA and Apoptin in the cells of groups A and B 48 and 72 h after transfection. The change in the ultrastructure of the cells in group A was observed under transmission electron microscope 72 h thereafter. MTT method was adopted to detect the cell proliferation activities of three groups 24,48,72 and 96 h thereafter and flow cytometry to determine the apoptosis rates and cell cycle changes 24,48 and 72 h thereafter. RESULTS:Compared to group B,group A demonstrated the expression of VP3 mRNA in SK-MES-1 and NCI-H1299 cell lines 48 h after transfection,and Apoptin expression and ultrastructure change for apopto-sis of SK-MES-1 and NCI-H1299 cell lines 72 h thereafter. Compared to groups B and C,group A showed lower proliferation activ-ities and higher apoptosis rates of SK-MES-1 and NCI-H1299 cell lines,which had a positive correlation with transfection time;and in the group A,there was a decrease in the proportion of the SK-MES-1 and NCI-H1299 cell lines in S phase and an increase in the proportion of those in G2/M phase,72 h after transfection. There was statistically difference (P<0.05). CONCLUSIONS:Adxsi-GFP-VP3 can effectively induce the apoptosis of SK-MES-1 and NCI-H1299 cell lines.
