Related Experiment Video
Updated: Aug 8, 2026
Using the E1A Minigene Tool to Study mRNA Splicing Changes
Published on: April 22, 2021
Effect of antisense MBD1 gene eukaryotic expression plasmid on expression of MBD1 gene in human biliary tract
Shi Zuo1, Shengquan Zou, Jian Luo
1Department of General Surgery, Tongji Hospital, Tongji Medical College, Huazhong Science and Technology University, Wuhan 430030, China.
Abstract:
Hypermethylation of the promoter region is one of the major mechanism of tumor suppressor gene inactivation. In order to provide a research tool for the study on the function of MBD1 gene in DNA methylation and tumorigenesis, antisense MBD1 gene eukaryotic expression plasmid was constructed and transfected into human biliary tract carcinoma cell line QBC-939 to observe its effect on the expression of MBD1 mRNA and protein by using RT-PCR and FCM respectively. Following the transfection, the mRNA level of MBD1 gene decreased from 0. 912 +/- 0.022 to 0.215 +/- 0. 017, and the protein level of MBD1 gene also decreased from (80.19 +/- 5.05) % to (35.11 +/- 4.05) %. There were very significant differences in the expression both at the transcription and post-transcription levels of MBD1 gene between non-tranfection group and the antisense MBD1 gene eukaryotic expression plasmid transfection group (P < 0.01). It was suggested that transfection with the antisense MBD1 gene eukaryotic expression plasmid can significantly reduce the expression level of MBD1 gene in QBC-939, and this study may provide a valid tool for the investigation of the function of MBD1 gene and its role in biliary tract carcinoma.
Insights
Researchers developed an antisense MBD1 gene plasmid to study its role in cancer. Transfection significantly reduced MBD1 gene expression in human biliary tract carcinoma cells, providing a tool for further research.
Area of Science:
- Molecular Biology
- Oncology
- Epigenetics
Background:
- Hypermethylation of promoter regions is a key mechanism for tumor suppressor gene inactivation.
- Understanding the role of Methyl CpG Binding Domain 1 (MBD1) gene in DNA methylation and tumorigenesis is crucial.
Purpose of the Study:
- To construct an antisense MBD1 gene eukaryotic expression plasmid.
- To investigate the effect of this plasmid on MBD1 gene expression in human biliary tract carcinoma cells (QBC-939).
- To provide a research tool for studying MBD1 gene function in tumorigenesis.
Main Methods:
- Construction and transfection of an antisense MBD1 gene eukaryotic expression plasmid into QBC-939 cells.
- Quantitative analysis of MBD1 mRNA levels using RT-PCR.
- Quantitative analysis of MBD1 protein levels using Flow Cytometry (FCM).
Main Results:
- Transfection significantly reduced MBD1 mRNA levels from 0.912 ± 0.022 to 0.215 ± 0.017.
- Transfection significantly reduced MBD1 protein levels from (80.19 ± 5.05)% to (35.11 ± 4.05)%.
- Statistically significant differences (P < 0.01) were observed at both transcriptional and post-transcriptional levels.
Conclusions:
- Antisense MBD1 gene plasmid transfection effectively reduces MBD1 gene expression in QBC-939 cells.
- This study provides a valuable tool for investigating MBD1 gene function and its role in biliary tract carcinoma.
