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TAp63 suppress metastasis via miR-133b in colon cancer cells.

Lin CW, Li XR, Zhang Y, Hu G, Guo YH, Zhou JY, Du J, Lv L, Gao K, Zhang Y, Deng H - Br. J. Cancer (2014)

Bottom Line: Furthermore, microRNA-133b is essential for the inhibitory effects of TAp63 on RhoA, E-cadherin and vimentin.Moreover, TAp63 inhibits cell migration and invasion through microRNA-133b.Correspondingly, the inhibitory effect of TAp63 on RhoA, E-cadherin, vimentin, migration and invasion can be blocked by the microRNA-133b inhibitor.

View Article: PubMed Central - PubMed

Affiliation: Department of General Surgery, The Third XiangYa Hospital of Central South University, Changsha, Hunan 410013, People's Republic of China.

ABSTRACT

Background: TAp63 is a tumour-suppressor protein that is often underexpressed in various types of cancer. It has been shown to activate gene transcription depending on the transcription domain and to be closely related with metastasis. In this study, we demonstrate that TAp63 suppresses metastasis in colon cancer cells through microRNA-133b.

Methods: We evaluated the correlation of TAp63 and miR-133b with HT-29 and SW-620 cells and investigated the roles of TAp63 in the expression of RhoA, E-cadherin and vimentin. We further investigated the roles of TAp63-mediated invasion and migration of colon cancer cells.

Results: TAp63 expression is downregulated in colon cancer, and microRNA-133b is a transcriptional target of TAp63. Furthermore, microRNA-133b is essential for the inhibitory effects of TAp63 on RhoA, E-cadherin and vimentin. Moreover, TAp63 inhibits cell migration and invasion through microRNA-133b. Correspondingly, the inhibitory effect of TAp63 on RhoA, E-cadherin, vimentin, migration and invasion can be blocked by the microRNA-133b inhibitor.

Conclusions: TAp63 and microRNA-133b were able to suppress the metastasis of colon cancer. Both TAp63 and microRNA-133b may be potential biomarkers for diagnosis in colon cancer metastasis and may provide unique therapeutic targets for this common malignancy.

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Expression of TAp63 in HT-29, SW-620 and CCD-18Co cells. The expression of TAp63 was analysed by RT–PCR, western blot and immunofluorescence. (A) Electrophoresis in 2% (w/v) agarose gel reveals a single 127-bp product following amplification by PCR using TAp63-specific primers. (B) Western blotting analysis of TAp63 on HT-29, SW-620 and CCD-18Co cells (top panel). The GAPDH loading control is shown in the bottom panel. (C) Immunofluorescence assay showed that TAp63 was present in the nuclei of HT-29, SW-620 and CCD-18Co cells. DAPI was used to counterstain the nucleus in blue, and TAp63 staining was in green. The figures show that the TAp63 expression level in HT-29 and SW-620 cells was much lower than that in CCD-18Co cells.
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fig1: Expression of TAp63 in HT-29, SW-620 and CCD-18Co cells. The expression of TAp63 was analysed by RT–PCR, western blot and immunofluorescence. (A) Electrophoresis in 2% (w/v) agarose gel reveals a single 127-bp product following amplification by PCR using TAp63-specific primers. (B) Western blotting analysis of TAp63 on HT-29, SW-620 and CCD-18Co cells (top panel). The GAPDH loading control is shown in the bottom panel. (C) Immunofluorescence assay showed that TAp63 was present in the nuclei of HT-29, SW-620 and CCD-18Co cells. DAPI was used to counterstain the nucleus in blue, and TAp63 staining was in green. The figures show that the TAp63 expression level in HT-29 and SW-620 cells was much lower than that in CCD-18Co cells.

Mentions: The TAp63 expression in HT-29, SW-620 and CCD-18Co cells was verified by RT–PCR, western blotting and immunofluorescence. The RT–PCR, western blotting and immunofluorescence positive results are shown in Figures 1A–C, respectively. The TAp63 expression level in HT-29 and SW-620 cells was much lower than that in CCD-18Co cells. Further, we analysed the expression of TAp63 in human colon cancer tissue and paired counterpart normal tissues using RT–PCR and western blotting. Lower levels of TAp63 were detected in the tumour tissues compared with the normal tissue (Figure 2).


TAp63 suppress metastasis via miR-133b in colon cancer cells.

Lin CW, Li XR, Zhang Y, Hu G, Guo YH, Zhou JY, Du J, Lv L, Gao K, Zhang Y, Deng H - Br. J. Cancer (2014)

Expression of TAp63 in HT-29, SW-620 and CCD-18Co cells. The expression of TAp63 was analysed by RT–PCR, western blot and immunofluorescence. (A) Electrophoresis in 2% (w/v) agarose gel reveals a single 127-bp product following amplification by PCR using TAp63-specific primers. (B) Western blotting analysis of TAp63 on HT-29, SW-620 and CCD-18Co cells (top panel). The GAPDH loading control is shown in the bottom panel. (C) Immunofluorescence assay showed that TAp63 was present in the nuclei of HT-29, SW-620 and CCD-18Co cells. DAPI was used to counterstain the nucleus in blue, and TAp63 staining was in green. The figures show that the TAp63 expression level in HT-29 and SW-620 cells was much lower than that in CCD-18Co cells.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4007221&req=5

fig1: Expression of TAp63 in HT-29, SW-620 and CCD-18Co cells. The expression of TAp63 was analysed by RT–PCR, western blot and immunofluorescence. (A) Electrophoresis in 2% (w/v) agarose gel reveals a single 127-bp product following amplification by PCR using TAp63-specific primers. (B) Western blotting analysis of TAp63 on HT-29, SW-620 and CCD-18Co cells (top panel). The GAPDH loading control is shown in the bottom panel. (C) Immunofluorescence assay showed that TAp63 was present in the nuclei of HT-29, SW-620 and CCD-18Co cells. DAPI was used to counterstain the nucleus in blue, and TAp63 staining was in green. The figures show that the TAp63 expression level in HT-29 and SW-620 cells was much lower than that in CCD-18Co cells.
Mentions: The TAp63 expression in HT-29, SW-620 and CCD-18Co cells was verified by RT–PCR, western blotting and immunofluorescence. The RT–PCR, western blotting and immunofluorescence positive results are shown in Figures 1A–C, respectively. The TAp63 expression level in HT-29 and SW-620 cells was much lower than that in CCD-18Co cells. Further, we analysed the expression of TAp63 in human colon cancer tissue and paired counterpart normal tissues using RT–PCR and western blotting. Lower levels of TAp63 were detected in the tumour tissues compared with the normal tissue (Figure 2).

Bottom Line: Furthermore, microRNA-133b is essential for the inhibitory effects of TAp63 on RhoA, E-cadherin and vimentin.Moreover, TAp63 inhibits cell migration and invasion through microRNA-133b.Correspondingly, the inhibitory effect of TAp63 on RhoA, E-cadherin, vimentin, migration and invasion can be blocked by the microRNA-133b inhibitor.

View Article: PubMed Central - PubMed

Affiliation: Department of General Surgery, The Third XiangYa Hospital of Central South University, Changsha, Hunan 410013, People's Republic of China.

ABSTRACT

Background: TAp63 is a tumour-suppressor protein that is often underexpressed in various types of cancer. It has been shown to activate gene transcription depending on the transcription domain and to be closely related with metastasis. In this study, we demonstrate that TAp63 suppresses metastasis in colon cancer cells through microRNA-133b.

Methods: We evaluated the correlation of TAp63 and miR-133b with HT-29 and SW-620 cells and investigated the roles of TAp63 in the expression of RhoA, E-cadherin and vimentin. We further investigated the roles of TAp63-mediated invasion and migration of colon cancer cells.

Results: TAp63 expression is downregulated in colon cancer, and microRNA-133b is a transcriptional target of TAp63. Furthermore, microRNA-133b is essential for the inhibitory effects of TAp63 on RhoA, E-cadherin and vimentin. Moreover, TAp63 inhibits cell migration and invasion through microRNA-133b. Correspondingly, the inhibitory effect of TAp63 on RhoA, E-cadherin, vimentin, migration and invasion can be blocked by the microRNA-133b inhibitor.

Conclusions: TAp63 and microRNA-133b were able to suppress the metastasis of colon cancer. Both TAp63 and microRNA-133b may be potential biomarkers for diagnosis in colon cancer metastasis and may provide unique therapeutic targets for this common malignancy.

Show MeSH
Related in: MedlinePlus