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The immunoglobulin heavy chain VH6-1 promoter regulates Ig transcription in non-B cells.

Wu L, Liu Y, Zhu X, Zhang L, Chen J, Zhang H, Hao P, Zhang S, Huang J, Zheng J, Zhang Y, Zhang Y, Qiu X - Cancer Cell Int. (2014)

Bottom Line: Our data showed that the sequence 1200 bp upstream of VH6-1 exhibited promoter activity in both B and non-B cells.No new regulatory elements were identified within the region 1200 bp to 300 bp upstream of VH6-1.The regulatory mechanisms among different cell types controlling the production of IgM heavy chains are worth discussing.

View Article: PubMed Central - PubMed

Affiliation: Key laboratory of Carcinogenesis and Translational Research (Ministry of Education), Central Laboratory, Peking University Cancer Hospital & Institute, Beijing, 100142 China.

ABSTRACT

Background: Non-B cell immunoglobulins (Igs) are widely expressed in epithelial cancer cells. The past 20 years of research have demonstrated that non-B cell Igs are associated with cancer cell proliferation, the cellular cytoskeleton and cancer stem cells. In this study we explored the transcriptional mechanism of IgM production in non-B cells.

Methods: The promoter region of a V-segment of the heavy mu chain gene (VH6-1) was cloned from a colon cancer cell line HT-29. Next, the promoter activities in non-B cells and B-cells were detected using the dual-luciferase reporter assay. Then the transcription factor binding to the promoter regions was evaluated by electrophoretic mobility shift assays (EMSAs) and gel supershift experiments.

Results: Our data showed that the sequence 1200 bp upstream of VH6-1 exhibited promoter activity in both B and non-B cells. No new regulatory elements were identified within the region 1200 bp to 300 bp upstream of VH6-1. In addition, Oct-1 was found to bind to the octamer element of the Ig gene promoter in cancer cells, in contrast to B cells, which utilize the transcriptional factor Oct-2.

Conclusion: The regulatory mechanisms among different cell types controlling the production of IgM heavy chains are worth discussing.

No MeSH data available.


Related in: MedlinePlus

The sequence 1200 bp to 300 bp upstream of VH6-1 contains no regulatory elements. (A) Schematic diagram of 5′-flanking 300-bp pGL3 construct. (B) The 300-bp deletion fragment amplified from the 1.2-kb fragment containing the VH6-1 promoter by PCR. (C) The 300-bp deletion truncations of the 1.2-kb pGL3 construct exhibited similar promoter activity to the 1.2-kb pGL3 construct in HT-29 and Raji cells.
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Fig3: The sequence 1200 bp to 300 bp upstream of VH6-1 contains no regulatory elements. (A) Schematic diagram of 5′-flanking 300-bp pGL3 construct. (B) The 300-bp deletion fragment amplified from the 1.2-kb fragment containing the VH6-1 promoter by PCR. (C) The 300-bp deletion truncations of the 1.2-kb pGL3 construct exhibited similar promoter activity to the 1.2-kb pGL3 construct in HT-29 and Raji cells.

Mentions: It was previously shown that the region 1200 bp to 300 bp upstream of VH4-59 contains two regulatory elements [15]. To determine whether regulatory elements exist in the sequence upstream of the VH6-1 promoter, the 1200-bp region containing the VH6-1 promoter was deleted via PCR (Figure 3A&B). The promoter activity of the deletion construct (pGL3-300 bp) was tested using a dual-luciferase reporter assay. Strong promoter activity was observed for both the 1200bb and 300-bp constructs in HT-29 and Raji cells, and the promoter activities of both were higher than for the SV40 promoter (Figure 3C). It is likely that there are no regulatory elements upstream (between 1200 bp and 300 bp) of the promoter of VH6-1 in cancer cells. This suggests that the mechanisms regulating the expression of VH6-1 and VH4-59 are different.Figure 3


The immunoglobulin heavy chain VH6-1 promoter regulates Ig transcription in non-B cells.

Wu L, Liu Y, Zhu X, Zhang L, Chen J, Zhang H, Hao P, Zhang S, Huang J, Zheng J, Zhang Y, Zhang Y, Qiu X - Cancer Cell Int. (2014)

The sequence 1200 bp to 300 bp upstream of VH6-1 contains no regulatory elements. (A) Schematic diagram of 5′-flanking 300-bp pGL3 construct. (B) The 300-bp deletion fragment amplified from the 1.2-kb fragment containing the VH6-1 promoter by PCR. (C) The 300-bp deletion truncations of the 1.2-kb pGL3 construct exhibited similar promoter activity to the 1.2-kb pGL3 construct in HT-29 and Raji cells.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4260249&req=5

Fig3: The sequence 1200 bp to 300 bp upstream of VH6-1 contains no regulatory elements. (A) Schematic diagram of 5′-flanking 300-bp pGL3 construct. (B) The 300-bp deletion fragment amplified from the 1.2-kb fragment containing the VH6-1 promoter by PCR. (C) The 300-bp deletion truncations of the 1.2-kb pGL3 construct exhibited similar promoter activity to the 1.2-kb pGL3 construct in HT-29 and Raji cells.
Mentions: It was previously shown that the region 1200 bp to 300 bp upstream of VH4-59 contains two regulatory elements [15]. To determine whether regulatory elements exist in the sequence upstream of the VH6-1 promoter, the 1200-bp region containing the VH6-1 promoter was deleted via PCR (Figure 3A&B). The promoter activity of the deletion construct (pGL3-300 bp) was tested using a dual-luciferase reporter assay. Strong promoter activity was observed for both the 1200bb and 300-bp constructs in HT-29 and Raji cells, and the promoter activities of both were higher than for the SV40 promoter (Figure 3C). It is likely that there are no regulatory elements upstream (between 1200 bp and 300 bp) of the promoter of VH6-1 in cancer cells. This suggests that the mechanisms regulating the expression of VH6-1 and VH4-59 are different.Figure 3

Bottom Line: Our data showed that the sequence 1200 bp upstream of VH6-1 exhibited promoter activity in both B and non-B cells.No new regulatory elements were identified within the region 1200 bp to 300 bp upstream of VH6-1.The regulatory mechanisms among different cell types controlling the production of IgM heavy chains are worth discussing.

View Article: PubMed Central - PubMed

Affiliation: Key laboratory of Carcinogenesis and Translational Research (Ministry of Education), Central Laboratory, Peking University Cancer Hospital & Institute, Beijing, 100142 China.

ABSTRACT

Background: Non-B cell immunoglobulins (Igs) are widely expressed in epithelial cancer cells. The past 20 years of research have demonstrated that non-B cell Igs are associated with cancer cell proliferation, the cellular cytoskeleton and cancer stem cells. In this study we explored the transcriptional mechanism of IgM production in non-B cells.

Methods: The promoter region of a V-segment of the heavy mu chain gene (VH6-1) was cloned from a colon cancer cell line HT-29. Next, the promoter activities in non-B cells and B-cells were detected using the dual-luciferase reporter assay. Then the transcription factor binding to the promoter regions was evaluated by electrophoretic mobility shift assays (EMSAs) and gel supershift experiments.

Results: Our data showed that the sequence 1200 bp upstream of VH6-1 exhibited promoter activity in both B and non-B cells. No new regulatory elements were identified within the region 1200 bp to 300 bp upstream of VH6-1. In addition, Oct-1 was found to bind to the octamer element of the Ig gene promoter in cancer cells, in contrast to B cells, which utilize the transcriptional factor Oct-2.

Conclusion: The regulatory mechanisms among different cell types controlling the production of IgM heavy chains are worth discussing.

No MeSH data available.


Related in: MedlinePlus