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Vasohibin-1 suppresses colon cancer.

Liu S, Han B, Zhang Q, Dou J, Wang F, Lin W, Sun Y, Peng G - Oncotarget (2015)

Bottom Line: Here we showed that stromal VASH1 levels were negatively correlated with tumor size, advanced clinical stage and distant metastases in colon cancer patients.Overexpression of VASH1 in colon cancer cells induced apoptosis and senescence, inhibiting cancer cell growth and colony formation in vitro and tumor growth in vivo.In addition, knockdown of VASH1 in cancer cells promoted cell growth, adhesion and migration in vitro, and enhanced tumorigenesis and metastasis in vivo.

View Article: PubMed Central - PubMed

Affiliation: Department of Oncology, Jinan Central Hospital, Affiliated to Shandong University, Jinan, P. R. China.

ABSTRACT
Vasohibin-1 (VASH1) is an endogenous angiogenesis inhibitor.However, the clinical relevance of VASH1 in colon cancer and its regulations on cancer angiogenesis and cancer cell biological characteristics are still unknown. Here we showed that stromal VASH1 levels were negatively correlated with tumor size, advanced clinical stage and distant metastases in colon cancer patients. Overexpression of VASH1 in colon cancer cells induced apoptosis and senescence, inhibiting cancer cell growth and colony formation in vitro and tumor growth in vivo. In addition, knockdown of VASH1 in cancer cells promoted cell growth, adhesion and migration in vitro, and enhanced tumorigenesis and metastasis in vivo.

No MeSH data available.


Related in: MedlinePlus

Knockdown of VASH1 in colon cancer HCT116 cells promotes cancer cell growth, adhesion and migration(A) The knockdown efficiency on VASH1-A and VASH1-B expression by the shRNA was determined in HCT116 tumor cells using Real-time PCR analyses. mRNA levels in each group were normalized to the relative quantity of GAPDH expression and compared against VASH1 expression level in control scramble shRNA group (set as 1). Results shown are mean ± SD from three independent experiments. **p<0.01 compared with the control scramble shRNA group. (B) and (C) Knockdown of VASH1 expression in HCT116 tumor cells dramatically promoted tumor cell growth and proliferation. HCT116 cells transfected with control shRNA served as a negative control. The cell growth of transfected HCT116 cells was evaluated at different time points using cell number counting (in B), and cell proliferation was determined using [3H]-thymidine assays (in C). **p<0.01 compared with the control scramble shRNA group. (D) Knockdown of VASH1 expression in HCT116 cells dramatically decreased the numbers and sizes of tumor colonies in colony formation assays. Results shown in the histogram are mean ± SD from three independent experiments. **p<0.01 compared with the control shRNA group. (E) Knockdown of VASH1 gene in HCT116 tumor cells markedly increased the adherent ability of tumor cells. The adhesion of transfected colon cancer cells was cultured in the fibronection-coated plates for 45 minutes. Adherent cells were counted and averaged in 10 fields at high (× 400) magnification with a microscope. **p<0.01 compared with the control shRNA group. (F) and (G) knockdown of VASH1 gene in HCT116 tumor cells significantly promoted the migration of tumor cells compared with the control shRNA-transfected tumor cells in the transwell migration assays (in F) and the wound closure assays (in G). *p<0.05 and **p<0.01 compared with the control shRNA group. Data in (A) to (G) are from three independent experiments with similar results.
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Figure 5: Knockdown of VASH1 in colon cancer HCT116 cells promotes cancer cell growth, adhesion and migration(A) The knockdown efficiency on VASH1-A and VASH1-B expression by the shRNA was determined in HCT116 tumor cells using Real-time PCR analyses. mRNA levels in each group were normalized to the relative quantity of GAPDH expression and compared against VASH1 expression level in control scramble shRNA group (set as 1). Results shown are mean ± SD from three independent experiments. **p<0.01 compared with the control scramble shRNA group. (B) and (C) Knockdown of VASH1 expression in HCT116 tumor cells dramatically promoted tumor cell growth and proliferation. HCT116 cells transfected with control shRNA served as a negative control. The cell growth of transfected HCT116 cells was evaluated at different time points using cell number counting (in B), and cell proliferation was determined using [3H]-thymidine assays (in C). **p<0.01 compared with the control scramble shRNA group. (D) Knockdown of VASH1 expression in HCT116 cells dramatically decreased the numbers and sizes of tumor colonies in colony formation assays. Results shown in the histogram are mean ± SD from three independent experiments. **p<0.01 compared with the control shRNA group. (E) Knockdown of VASH1 gene in HCT116 tumor cells markedly increased the adherent ability of tumor cells. The adhesion of transfected colon cancer cells was cultured in the fibronection-coated plates for 45 minutes. Adherent cells were counted and averaged in 10 fields at high (× 400) magnification with a microscope. **p<0.01 compared with the control shRNA group. (F) and (G) knockdown of VASH1 gene in HCT116 tumor cells significantly promoted the migration of tumor cells compared with the control shRNA-transfected tumor cells in the transwell migration assays (in F) and the wound closure assays (in G). *p<0.05 and **p<0.01 compared with the control shRNA group. Data in (A) to (G) are from three independent experiments with similar results.

Mentions: To further confirm the functional role of VASH1 in regulating colon cancer cell growth, we also utilized the loss-of-function strategy to knockdown VASH1 gene with shRNA in VASH1 highly expressed HCT116 tumor cells and then determined its effect on tumor growth and proliferation. We selected a shRNA which can specifically target both VASH1-A and VASH1-B isoforms (Supplemental Figure 4A). The knockdown efficiency on VASH1 expression by the shRNA was further confirmed in both VASH1-transfected 293T cells and in HCT116 tumor cells, using western-blot and Real-time PCR analyses, respectively (Supplemental Figure 4B and 4C, and Figure 5A). As expected, silence of VASH1 expression in HCT116 tumor cells dramatically promoted tumor growth and increased cell proliferation (Figure 5B and 5C). Furthermore, the numbers and sizes of tumor cell colonies were also significantly increased in HCT116 cells after knockdown of VASH1 gene in a colony formation assay (Figure 5D). These results further suggested that VASH1 expression in colon cancer cells directly controlled cell growth and fate.


Vasohibin-1 suppresses colon cancer.

Liu S, Han B, Zhang Q, Dou J, Wang F, Lin W, Sun Y, Peng G - Oncotarget (2015)

Knockdown of VASH1 in colon cancer HCT116 cells promotes cancer cell growth, adhesion and migration(A) The knockdown efficiency on VASH1-A and VASH1-B expression by the shRNA was determined in HCT116 tumor cells using Real-time PCR analyses. mRNA levels in each group were normalized to the relative quantity of GAPDH expression and compared against VASH1 expression level in control scramble shRNA group (set as 1). Results shown are mean ± SD from three independent experiments. **p<0.01 compared with the control scramble shRNA group. (B) and (C) Knockdown of VASH1 expression in HCT116 tumor cells dramatically promoted tumor cell growth and proliferation. HCT116 cells transfected with control shRNA served as a negative control. The cell growth of transfected HCT116 cells was evaluated at different time points using cell number counting (in B), and cell proliferation was determined using [3H]-thymidine assays (in C). **p<0.01 compared with the control scramble shRNA group. (D) Knockdown of VASH1 expression in HCT116 cells dramatically decreased the numbers and sizes of tumor colonies in colony formation assays. Results shown in the histogram are mean ± SD from three independent experiments. **p<0.01 compared with the control shRNA group. (E) Knockdown of VASH1 gene in HCT116 tumor cells markedly increased the adherent ability of tumor cells. The adhesion of transfected colon cancer cells was cultured in the fibronection-coated plates for 45 minutes. Adherent cells were counted and averaged in 10 fields at high (× 400) magnification with a microscope. **p<0.01 compared with the control shRNA group. (F) and (G) knockdown of VASH1 gene in HCT116 tumor cells significantly promoted the migration of tumor cells compared with the control shRNA-transfected tumor cells in the transwell migration assays (in F) and the wound closure assays (in G). *p<0.05 and **p<0.01 compared with the control shRNA group. Data in (A) to (G) are from three independent experiments with similar results.
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Related In: Results  -  Collection

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Figure 5: Knockdown of VASH1 in colon cancer HCT116 cells promotes cancer cell growth, adhesion and migration(A) The knockdown efficiency on VASH1-A and VASH1-B expression by the shRNA was determined in HCT116 tumor cells using Real-time PCR analyses. mRNA levels in each group were normalized to the relative quantity of GAPDH expression and compared against VASH1 expression level in control scramble shRNA group (set as 1). Results shown are mean ± SD from three independent experiments. **p<0.01 compared with the control scramble shRNA group. (B) and (C) Knockdown of VASH1 expression in HCT116 tumor cells dramatically promoted tumor cell growth and proliferation. HCT116 cells transfected with control shRNA served as a negative control. The cell growth of transfected HCT116 cells was evaluated at different time points using cell number counting (in B), and cell proliferation was determined using [3H]-thymidine assays (in C). **p<0.01 compared with the control scramble shRNA group. (D) Knockdown of VASH1 expression in HCT116 cells dramatically decreased the numbers and sizes of tumor colonies in colony formation assays. Results shown in the histogram are mean ± SD from three independent experiments. **p<0.01 compared with the control shRNA group. (E) Knockdown of VASH1 gene in HCT116 tumor cells markedly increased the adherent ability of tumor cells. The adhesion of transfected colon cancer cells was cultured in the fibronection-coated plates for 45 minutes. Adherent cells were counted and averaged in 10 fields at high (× 400) magnification with a microscope. **p<0.01 compared with the control shRNA group. (F) and (G) knockdown of VASH1 gene in HCT116 tumor cells significantly promoted the migration of tumor cells compared with the control shRNA-transfected tumor cells in the transwell migration assays (in F) and the wound closure assays (in G). *p<0.05 and **p<0.01 compared with the control shRNA group. Data in (A) to (G) are from three independent experiments with similar results.
Mentions: To further confirm the functional role of VASH1 in regulating colon cancer cell growth, we also utilized the loss-of-function strategy to knockdown VASH1 gene with shRNA in VASH1 highly expressed HCT116 tumor cells and then determined its effect on tumor growth and proliferation. We selected a shRNA which can specifically target both VASH1-A and VASH1-B isoforms (Supplemental Figure 4A). The knockdown efficiency on VASH1 expression by the shRNA was further confirmed in both VASH1-transfected 293T cells and in HCT116 tumor cells, using western-blot and Real-time PCR analyses, respectively (Supplemental Figure 4B and 4C, and Figure 5A). As expected, silence of VASH1 expression in HCT116 tumor cells dramatically promoted tumor growth and increased cell proliferation (Figure 5B and 5C). Furthermore, the numbers and sizes of tumor cell colonies were also significantly increased in HCT116 cells after knockdown of VASH1 gene in a colony formation assay (Figure 5D). These results further suggested that VASH1 expression in colon cancer cells directly controlled cell growth and fate.

Bottom Line: Here we showed that stromal VASH1 levels were negatively correlated with tumor size, advanced clinical stage and distant metastases in colon cancer patients.Overexpression of VASH1 in colon cancer cells induced apoptosis and senescence, inhibiting cancer cell growth and colony formation in vitro and tumor growth in vivo.In addition, knockdown of VASH1 in cancer cells promoted cell growth, adhesion and migration in vitro, and enhanced tumorigenesis and metastasis in vivo.

View Article: PubMed Central - PubMed

Affiliation: Department of Oncology, Jinan Central Hospital, Affiliated to Shandong University, Jinan, P. R. China.

ABSTRACT
Vasohibin-1 (VASH1) is an endogenous angiogenesis inhibitor.However, the clinical relevance of VASH1 in colon cancer and its regulations on cancer angiogenesis and cancer cell biological characteristics are still unknown. Here we showed that stromal VASH1 levels were negatively correlated with tumor size, advanced clinical stage and distant metastases in colon cancer patients. Overexpression of VASH1 in colon cancer cells induced apoptosis and senescence, inhibiting cancer cell growth and colony formation in vitro and tumor growth in vivo. In addition, knockdown of VASH1 in cancer cells promoted cell growth, adhesion and migration in vitro, and enhanced tumorigenesis and metastasis in vivo.

No MeSH data available.


Related in: MedlinePlus