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Combining TGF-β1 knockdown and miR200c administration to optimize antitumor efficacy of B16F10/GPI-IL-21 vaccine.

Wang X, Zhao F, He X, Wang J, Zhang Y, Zhang H, Ni Y, Sun J, Wang X, Dou J - Oncotarget (2015)

Bottom Line: Our study aimed at optimizing the antitumor effects of the B16F10/glycosylphosphatidylinositol-interleukin 21 (B16F10/GPI-IL-21) tumor vaccine on melanoma bearing mice by combining the TGF-β1 knockdown and the administration of miR200c agomir.The later combination showed that, when compared with the mice in the control group that received no vaccination, vaccinated mice significantly increased NK and CTL activities, enhanced levels of IFN-γ, and reduced expression of TGF-β1, N-cadherin, Vimentin, Gli1/2, P-Smad2/3 and others involved in promoting expression of EMT-related molecules in tumor areas, and inhibited the melanoma metastasis in lungs and lymph nodes.Altogether, our findings demonstrate that this synergistic anti-cancer regimen effectively induces strong immune response and diminishes the melanoma progression.

View Article: PubMed Central - PubMed

Affiliation: Department of Pathogenic Biology and Immunology, School of Medicine & Collaborative Innovation Center of Suzhou Nano Science and Technology, Southeast University, Nanjing, China.

ABSTRACT
TGF-β1 secreted abundantly by tumors cells as well as present in the local microenvironment promotes neoplasm invasion and metastasis by triggering the epithelial to mesenchymal transition (EMT). MiR200c has been shown to suppress EMT and to regulate the cellular epithelial and interstitial state conversion, whereas the tumor vaccines are intended to specifically initiate or amplify a host response against evolving tumor cells. Our study aimed at optimizing the antitumor effects of the B16F10/glycosylphosphatidylinositol-interleukin 21 (B16F10/GPI-IL-21) tumor vaccine on melanoma bearing mice by combining the TGF-β1 knockdown and the administration of miR200c agomir. The mice were subcutaneously vaccinated with inactivated B16F10/GPI-IL-21 vaccine and challenged by B16F10 cells transfected with shTGF-β1 (B16F10/shTGF-β1 cells) or B16F10/shTGF-β1 cells with the administration of miR200c agomir. The later combination showed that, when compared with the mice in the control group that received no vaccination, vaccinated mice significantly increased NK and CTL activities, enhanced levels of IFN-γ, and reduced expression of TGF-β1, N-cadherin, Vimentin, Gli1/2, P-Smad2/3 and others involved in promoting expression of EMT-related molecules in tumor areas, and inhibited the melanoma metastasis in lungs and lymph nodes. Altogether, our findings demonstrate that this synergistic anti-cancer regimen effectively induces strong immune response and diminishes the melanoma progression.

No MeSH data available.


Related in: MedlinePlus

Tumor growth and metastasis in vaccinated mice challenged with B16F10/shTGF-β1 cells plus minus miR200c agomirA. Images shows the tumor sizes dissected from the differently treated mice on Day 51. After the mice were initially immunized s.c. with 1×106 B16F10/GPI-IL-21 inactivated tumor vaccine the mice were challenged by the differently treated B16F10 cells as described in the section of materials and methods. B. The quantification analysis of tumor sizes (*p < 0.05, ** p < 0.01 and *** p < 0.001). C. Tumor free mice during 51 days. D. Images of tumor metastatic focus in mouse lungs. E. The quantification analysis of lung metastatic focus counts (HPF, ×400). Data are represented as mean +/− SEM; refer to the statistical differences as indicated.
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Figure 5: Tumor growth and metastasis in vaccinated mice challenged with B16F10/shTGF-β1 cells plus minus miR200c agomirA. Images shows the tumor sizes dissected from the differently treated mice on Day 51. After the mice were initially immunized s.c. with 1×106 B16F10/GPI-IL-21 inactivated tumor vaccine the mice were challenged by the differently treated B16F10 cells as described in the section of materials and methods. B. The quantification analysis of tumor sizes (*p < 0.05, ** p < 0.01 and *** p < 0.001). C. Tumor free mice during 51 days. D. Images of tumor metastatic focus in mouse lungs. E. The quantification analysis of lung metastatic focus counts (HPF, ×400). Data are represented as mean +/− SEM; refer to the statistical differences as indicated.

Mentions: To reinforce the antitumor efficacy of B16F10/GPI-IL-21 in the B16F10 melanoma-bearing C57BL/6 mice, we down regulated the TGF-β1 expression in the B16F10 cells by the RNAi technology and injection of miR200c agomir simultaneously. Figure 5A portrays the sizes of tumor dissected from the mice challenged by the differently treated B16F10 cells. We found that 3 of the 6 mice developed tumors on Day 18, Day 21 and Day 24, respectively after the mice were challenged by the B16F10-shTGF-β1 cells, and that 2 of the 6 mice challenged by the B16F10-shTGF-β1 cells with the injection of miR200c agomir developed tumors on Day 21 and Day 27, respectively (Figure 5B). Figure 5A-5B also show that the B16F10/GPI-IL-21 immunized mice developed tumors on Day 24 and Day 27, respectively, after the 6 mice were challenged by the B16F10-shTGF-β1 cells. In contrast, only 1 of the 6 mice in the experiment group grew the smallest tumor on Day 33 (Figure 5A-5C) along with the obvious reduction in pulmonary metastasis (Figure 5D) in the group challenged by B16F10/GPI-IL-21 combined with B16F10-shTGF-β1 and miR200c. No measurable tumors were detected in the remaining 5 mice until 51 days into the observation (Figure 5C).


Combining TGF-β1 knockdown and miR200c administration to optimize antitumor efficacy of B16F10/GPI-IL-21 vaccine.

Wang X, Zhao F, He X, Wang J, Zhang Y, Zhang H, Ni Y, Sun J, Wang X, Dou J - Oncotarget (2015)

Tumor growth and metastasis in vaccinated mice challenged with B16F10/shTGF-β1 cells plus minus miR200c agomirA. Images shows the tumor sizes dissected from the differently treated mice on Day 51. After the mice were initially immunized s.c. with 1×106 B16F10/GPI-IL-21 inactivated tumor vaccine the mice were challenged by the differently treated B16F10 cells as described in the section of materials and methods. B. The quantification analysis of tumor sizes (*p < 0.05, ** p < 0.01 and *** p < 0.001). C. Tumor free mice during 51 days. D. Images of tumor metastatic focus in mouse lungs. E. The quantification analysis of lung metastatic focus counts (HPF, ×400). Data are represented as mean +/− SEM; refer to the statistical differences as indicated.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4494953&req=5

Figure 5: Tumor growth and metastasis in vaccinated mice challenged with B16F10/shTGF-β1 cells plus minus miR200c agomirA. Images shows the tumor sizes dissected from the differently treated mice on Day 51. After the mice were initially immunized s.c. with 1×106 B16F10/GPI-IL-21 inactivated tumor vaccine the mice were challenged by the differently treated B16F10 cells as described in the section of materials and methods. B. The quantification analysis of tumor sizes (*p < 0.05, ** p < 0.01 and *** p < 0.001). C. Tumor free mice during 51 days. D. Images of tumor metastatic focus in mouse lungs. E. The quantification analysis of lung metastatic focus counts (HPF, ×400). Data are represented as mean +/− SEM; refer to the statistical differences as indicated.
Mentions: To reinforce the antitumor efficacy of B16F10/GPI-IL-21 in the B16F10 melanoma-bearing C57BL/6 mice, we down regulated the TGF-β1 expression in the B16F10 cells by the RNAi technology and injection of miR200c agomir simultaneously. Figure 5A portrays the sizes of tumor dissected from the mice challenged by the differently treated B16F10 cells. We found that 3 of the 6 mice developed tumors on Day 18, Day 21 and Day 24, respectively after the mice were challenged by the B16F10-shTGF-β1 cells, and that 2 of the 6 mice challenged by the B16F10-shTGF-β1 cells with the injection of miR200c agomir developed tumors on Day 21 and Day 27, respectively (Figure 5B). Figure 5A-5B also show that the B16F10/GPI-IL-21 immunized mice developed tumors on Day 24 and Day 27, respectively, after the 6 mice were challenged by the B16F10-shTGF-β1 cells. In contrast, only 1 of the 6 mice in the experiment group grew the smallest tumor on Day 33 (Figure 5A-5C) along with the obvious reduction in pulmonary metastasis (Figure 5D) in the group challenged by B16F10/GPI-IL-21 combined with B16F10-shTGF-β1 and miR200c. No measurable tumors were detected in the remaining 5 mice until 51 days into the observation (Figure 5C).

Bottom Line: Our study aimed at optimizing the antitumor effects of the B16F10/glycosylphosphatidylinositol-interleukin 21 (B16F10/GPI-IL-21) tumor vaccine on melanoma bearing mice by combining the TGF-β1 knockdown and the administration of miR200c agomir.The later combination showed that, when compared with the mice in the control group that received no vaccination, vaccinated mice significantly increased NK and CTL activities, enhanced levels of IFN-γ, and reduced expression of TGF-β1, N-cadherin, Vimentin, Gli1/2, P-Smad2/3 and others involved in promoting expression of EMT-related molecules in tumor areas, and inhibited the melanoma metastasis in lungs and lymph nodes.Altogether, our findings demonstrate that this synergistic anti-cancer regimen effectively induces strong immune response and diminishes the melanoma progression.

View Article: PubMed Central - PubMed

Affiliation: Department of Pathogenic Biology and Immunology, School of Medicine & Collaborative Innovation Center of Suzhou Nano Science and Technology, Southeast University, Nanjing, China.

ABSTRACT
TGF-β1 secreted abundantly by tumors cells as well as present in the local microenvironment promotes neoplasm invasion and metastasis by triggering the epithelial to mesenchymal transition (EMT). MiR200c has been shown to suppress EMT and to regulate the cellular epithelial and interstitial state conversion, whereas the tumor vaccines are intended to specifically initiate or amplify a host response against evolving tumor cells. Our study aimed at optimizing the antitumor effects of the B16F10/glycosylphosphatidylinositol-interleukin 21 (B16F10/GPI-IL-21) tumor vaccine on melanoma bearing mice by combining the TGF-β1 knockdown and the administration of miR200c agomir. The mice were subcutaneously vaccinated with inactivated B16F10/GPI-IL-21 vaccine and challenged by B16F10 cells transfected with shTGF-β1 (B16F10/shTGF-β1 cells) or B16F10/shTGF-β1 cells with the administration of miR200c agomir. The later combination showed that, when compared with the mice in the control group that received no vaccination, vaccinated mice significantly increased NK and CTL activities, enhanced levels of IFN-γ, and reduced expression of TGF-β1, N-cadherin, Vimentin, Gli1/2, P-Smad2/3 and others involved in promoting expression of EMT-related molecules in tumor areas, and inhibited the melanoma metastasis in lungs and lymph nodes. Altogether, our findings demonstrate that this synergistic anti-cancer regimen effectively induces strong immune response and diminishes the melanoma progression.

No MeSH data available.


Related in: MedlinePlus