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Construction of interference vector targeting Ep-CAM gene and its effects on colorectal cancer cell proliferation.

Qi Y, Zhou F, Zhang L, Liu L, Xu H, Guo H - Drug Des Devel Ther (2015)

Bottom Line: The results indicated that the Ep-CAM messenger (m)RNA expression in the interference group was lower significantly compared with that of the empty plasmid group and control group (P<0.01).Western blot analysis results showed that Ep-CAM protein expression was significantly lower in interference group compared with that of the empty plasmid group and the control group (P<0.01).MTT assay results demonstrated that the proliferation ability of cells in the interference group was significantly inhibited compared with the two other groups (P<0.05).

View Article: PubMed Central - PubMed

Affiliation: Department of Gastroenterology, Binzhou People's Hospital, Binzhou, Shandong, People's Republic of China.

ABSTRACT

Background: Prior study indicates that abnormal protein expression and functional changes in the development and progression of colorectal cancer is related to gene expression. The aim of this study was to construct an interference plasmid targeting the Ep-CAM gene and to investigate its effects on the proliferation of colorectal cancer cells.

Methods: In this study, HT-29 and HCT-116 colorectal cancer cell lines were selected as cell models. The double-stranded micro (mi)RNA oligo was inserted into the pcDNATM6.2-GW/EmGFPmiR vector, which is an expression of miRNA. Lipofectamine™ 2000 was used to transfer plasmid into the empty plasmid group (transfected pcDNATM6.2-GW/EmGFPmiR-neg) and the interference group (transfected pcDNATM6.2-GW/EmGFPmiR-Ep-CAM-1), respectively. Meanwhile, the nontransferred HT-29 and HCT-116 acts as the blank control group. Reverse transcription polymerase chain reaction (RT-PCR) was used to detect the transfection efficiency. Western blot was used to detect Ep-CAM protein expression. The cell proliferation in each group was detected by using 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay.

Results: The results indicated that the Ep-CAM messenger (m)RNA expression in the interference group was lower significantly compared with that of the empty plasmid group and control group (P<0.01). Western blot analysis results showed that Ep-CAM protein expression was significantly lower in interference group compared with that of the empty plasmid group and the control group (P<0.01). MTT assay results demonstrated that the proliferation ability of cells in the interference group was significantly inhibited compared with the two other groups (P<0.05).

Conclusion: Silencing of Ep-CAM can significantly inhibit the proliferation of colorectal cancer cells.

No MeSH data available.


Related in: MedlinePlus

Detection of expression of Ep-CAM protein with western blot.Notes: (A) Ep-CAM expression in HT-29 cells. (B) Ep-CAM expression in HCT-116 cells. Western blot results showed that in HT-29 and HCT-116 cells, expression of Ep-CAM protein was significantly lower in the interference group than in the empty plasmid group and blank control group.
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f2-dddt-9-2647: Detection of expression of Ep-CAM protein with western blot.Notes: (A) Ep-CAM expression in HT-29 cells. (B) Ep-CAM expression in HCT-116 cells. Western blot results showed that in HT-29 and HCT-116 cells, expression of Ep-CAM protein was significantly lower in the interference group than in the empty plasmid group and blank control group.

Mentions: The Ep-CAM protein expression in HT-29 and HCT-116 cell lines was also examined using the western blot assay. The results showed that the Ep-CAM expression in the interference group was significantly lower compared with that of the empty vector group and blank control group (Figure 2) (P<0.05). Furthermore, there was no significant difference for Ep-CAM expression between the empty plasmid group and blank control group (Figure 2) (P<0.05).


Construction of interference vector targeting Ep-CAM gene and its effects on colorectal cancer cell proliferation.

Qi Y, Zhou F, Zhang L, Liu L, Xu H, Guo H - Drug Des Devel Ther (2015)

Detection of expression of Ep-CAM protein with western blot.Notes: (A) Ep-CAM expression in HT-29 cells. (B) Ep-CAM expression in HCT-116 cells. Western blot results showed that in HT-29 and HCT-116 cells, expression of Ep-CAM protein was significantly lower in the interference group than in the empty plasmid group and blank control group.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4440878&req=5

f2-dddt-9-2647: Detection of expression of Ep-CAM protein with western blot.Notes: (A) Ep-CAM expression in HT-29 cells. (B) Ep-CAM expression in HCT-116 cells. Western blot results showed that in HT-29 and HCT-116 cells, expression of Ep-CAM protein was significantly lower in the interference group than in the empty plasmid group and blank control group.
Mentions: The Ep-CAM protein expression in HT-29 and HCT-116 cell lines was also examined using the western blot assay. The results showed that the Ep-CAM expression in the interference group was significantly lower compared with that of the empty vector group and blank control group (Figure 2) (P<0.05). Furthermore, there was no significant difference for Ep-CAM expression between the empty plasmid group and blank control group (Figure 2) (P<0.05).

Bottom Line: The results indicated that the Ep-CAM messenger (m)RNA expression in the interference group was lower significantly compared with that of the empty plasmid group and control group (P<0.01).Western blot analysis results showed that Ep-CAM protein expression was significantly lower in interference group compared with that of the empty plasmid group and the control group (P<0.01).MTT assay results demonstrated that the proliferation ability of cells in the interference group was significantly inhibited compared with the two other groups (P<0.05).

View Article: PubMed Central - PubMed

Affiliation: Department of Gastroenterology, Binzhou People's Hospital, Binzhou, Shandong, People's Republic of China.

ABSTRACT

Background: Prior study indicates that abnormal protein expression and functional changes in the development and progression of colorectal cancer is related to gene expression. The aim of this study was to construct an interference plasmid targeting the Ep-CAM gene and to investigate its effects on the proliferation of colorectal cancer cells.

Methods: In this study, HT-29 and HCT-116 colorectal cancer cell lines were selected as cell models. The double-stranded micro (mi)RNA oligo was inserted into the pcDNATM6.2-GW/EmGFPmiR vector, which is an expression of miRNA. Lipofectamine™ 2000 was used to transfer plasmid into the empty plasmid group (transfected pcDNATM6.2-GW/EmGFPmiR-neg) and the interference group (transfected pcDNATM6.2-GW/EmGFPmiR-Ep-CAM-1), respectively. Meanwhile, the nontransferred HT-29 and HCT-116 acts as the blank control group. Reverse transcription polymerase chain reaction (RT-PCR) was used to detect the transfection efficiency. Western blot was used to detect Ep-CAM protein expression. The cell proliferation in each group was detected by using 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay.

Results: The results indicated that the Ep-CAM messenger (m)RNA expression in the interference group was lower significantly compared with that of the empty plasmid group and control group (P<0.01). Western blot analysis results showed that Ep-CAM protein expression was significantly lower in interference group compared with that of the empty plasmid group and the control group (P<0.01). MTT assay results demonstrated that the proliferation ability of cells in the interference group was significantly inhibited compared with the two other groups (P<0.05).

Conclusion: Silencing of Ep-CAM can significantly inhibit the proliferation of colorectal cancer cells.

No MeSH data available.


Related in: MedlinePlus