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Anti-IGF-1R monoclonal antibody inhibits the carcinogenicity activity of acquired trastuzumab-resistant SKOV3.

Wang W, Zhang Y, Lv M, Feng J, Peng H, Geng J, Lin Z, Zhou T, Li X, Shen B, Ma Y, Qiao C - J Ovarian Res (2014)

Bottom Line: Reversing the resistance often results in better clinical therapeutic effect.It was also confirmed preliminarily that the mechanism of antibody might be to inhibit the activation of IGF-1R and downstream MAPK, AKT pathway transduction.In similar cases, not only acquired but also de novo, good curative effect might be achieved using combined antibody therapy strategies.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Cellular and Molecular Immunology, Institute of Immunology, Henan University, Kaifeng, 475001, China. 1790735517@qq.com.

ABSTRACT

Background: Antibody resistance, not only de novo but also acquired cases, usually exists and is related with lower survival rate and high risk of recurrence. Reversing the resistance often results in better clinical therapeutic effect. Previously, we established a trastuzumab-resistant ovarian cancer cell line, named as SKOV3-T, with lower HER2 and induced higher IGF-1R expression level to keep cell survival.

Methods: IGF-1R was identified important for SKOV3-T growth. Then, a novel anti-IGF-1R monoclonal antibody, named as LMAb1, was used to inhibit SKOV3-T in cell growth/proliferation, migration, clone formation and in vivo carcinogenicity.

Results: In both in vitro and in vivo assays, LMAb1 showed effective anti-tumor function, especially when being used in combination with trastuzumab, which was beneficial to longer survival time of mice as well as smaller tumor. It was also confirmed preliminarily that the mechanism of antibody might be to inhibit the activation of IGF-1R and downstream MAPK, AKT pathway transduction.

Conclusion: We achieved satisfactory anti-tumor activity using trastuzumab plus LMAb1 in trastuzumab-resistant ovarian cancer model. In similar cases, not only acquired but also de novo, good curative effect might be achieved using combined antibody therapy strategies.

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Related in: MedlinePlus

IGF-1R can promote the proliferation of SKOV3. (A) IGF-1R expression was up-regulated in SKOV3-T while HER2 was opposite by flow cytometry analysis; (B) IGF-1R-positive SKOV3 cell preparation by eukaryotic transfection with pCMV6-IGF-1R plasmid. Cell proliferation (C) and agar clone formation (D) assays both indicated the enhanced carcinogenic activity of IGF-1R-positive SKOV3 cells, while according to cell cycle analysis (E), SKOV3-IGF1R owned more S-phase cells in order to multiply quicker; Similarly, in vivo tumor model (F) further displayed more rapid tumor growth of SKOV3-IGF1R, indicating that IGF-1R can promote the cell survival and multiplication in ovarian cancer SKOV3 cells. To be clear, in this figure, “SKOV3” sample means original pCMV6 transfected cells.
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Fig2: IGF-1R can promote the proliferation of SKOV3. (A) IGF-1R expression was up-regulated in SKOV3-T while HER2 was opposite by flow cytometry analysis; (B) IGF-1R-positive SKOV3 cell preparation by eukaryotic transfection with pCMV6-IGF-1R plasmid. Cell proliferation (C) and agar clone formation (D) assays both indicated the enhanced carcinogenic activity of IGF-1R-positive SKOV3 cells, while according to cell cycle analysis (E), SKOV3-IGF1R owned more S-phase cells in order to multiply quicker; Similarly, in vivo tumor model (F) further displayed more rapid tumor growth of SKOV3-IGF1R, indicating that IGF-1R can promote the cell survival and multiplication in ovarian cancer SKOV3 cells. To be clear, in this figure, “SKOV3” sample means original pCMV6 transfected cells.

Mentions: According to our previous work, based on the mRNA array analysis, IGF-1R up-regulation (Figure 2A) was proved to play a key role in SKOV3-T proliferation in vitro. To the opposite, the expression level of HER2 in SKOV3-T was dramatically lower than SKOV3by flow cytometry method (Figure 2A).To further examine the role of IGF-1R in ovarian cancer cells, IGF-1R-pCMV6 plasmid, a eukaryotic expression vector subcloned with full IGF-1R exon sequence, was transfected into SKOV3 using lipofectamine 2000, generating a pool of IGF-1R-positive SKOV3 cells (Figure 2B). As shown in Figure 2C, the transfected cells had much higher viability or proliferative capacity than SKOV3; meanwhile, they also displayed increased colony formation capacity (Figure 2D); furthermore, according to the cell cycling assay, SKOV3-IGF-1R cells (pool) has S-phase cells than SKOV3, suggesting much quicker cell multiplication rate of SKOV3-IGF-1Rcells (Figure 2E); in in vivo tumor formation assay, IGF-1R positive cells exhibited more rapid growth capacity than parental cells (Figure 2F), indicating thatIGF-1R can promote the proliferation of SKOV3. All above demonstrated the importance of membrane IGF-1R as well as its downstream cascade in retaining/promoting the survival of SKOV3-T, especially when HER2-related signal pathway was down-regulated.Figure 2


Anti-IGF-1R monoclonal antibody inhibits the carcinogenicity activity of acquired trastuzumab-resistant SKOV3.

Wang W, Zhang Y, Lv M, Feng J, Peng H, Geng J, Lin Z, Zhou T, Li X, Shen B, Ma Y, Qiao C - J Ovarian Res (2014)

IGF-1R can promote the proliferation of SKOV3. (A) IGF-1R expression was up-regulated in SKOV3-T while HER2 was opposite by flow cytometry analysis; (B) IGF-1R-positive SKOV3 cell preparation by eukaryotic transfection with pCMV6-IGF-1R plasmid. Cell proliferation (C) and agar clone formation (D) assays both indicated the enhanced carcinogenic activity of IGF-1R-positive SKOV3 cells, while according to cell cycle analysis (E), SKOV3-IGF1R owned more S-phase cells in order to multiply quicker; Similarly, in vivo tumor model (F) further displayed more rapid tumor growth of SKOV3-IGF1R, indicating that IGF-1R can promote the cell survival and multiplication in ovarian cancer SKOV3 cells. To be clear, in this figure, “SKOV3” sample means original pCMV6 transfected cells.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4260252&req=5

Fig2: IGF-1R can promote the proliferation of SKOV3. (A) IGF-1R expression was up-regulated in SKOV3-T while HER2 was opposite by flow cytometry analysis; (B) IGF-1R-positive SKOV3 cell preparation by eukaryotic transfection with pCMV6-IGF-1R plasmid. Cell proliferation (C) and agar clone formation (D) assays both indicated the enhanced carcinogenic activity of IGF-1R-positive SKOV3 cells, while according to cell cycle analysis (E), SKOV3-IGF1R owned more S-phase cells in order to multiply quicker; Similarly, in vivo tumor model (F) further displayed more rapid tumor growth of SKOV3-IGF1R, indicating that IGF-1R can promote the cell survival and multiplication in ovarian cancer SKOV3 cells. To be clear, in this figure, “SKOV3” sample means original pCMV6 transfected cells.
Mentions: According to our previous work, based on the mRNA array analysis, IGF-1R up-regulation (Figure 2A) was proved to play a key role in SKOV3-T proliferation in vitro. To the opposite, the expression level of HER2 in SKOV3-T was dramatically lower than SKOV3by flow cytometry method (Figure 2A).To further examine the role of IGF-1R in ovarian cancer cells, IGF-1R-pCMV6 plasmid, a eukaryotic expression vector subcloned with full IGF-1R exon sequence, was transfected into SKOV3 using lipofectamine 2000, generating a pool of IGF-1R-positive SKOV3 cells (Figure 2B). As shown in Figure 2C, the transfected cells had much higher viability or proliferative capacity than SKOV3; meanwhile, they also displayed increased colony formation capacity (Figure 2D); furthermore, according to the cell cycling assay, SKOV3-IGF-1R cells (pool) has S-phase cells than SKOV3, suggesting much quicker cell multiplication rate of SKOV3-IGF-1Rcells (Figure 2E); in in vivo tumor formation assay, IGF-1R positive cells exhibited more rapid growth capacity than parental cells (Figure 2F), indicating thatIGF-1R can promote the proliferation of SKOV3. All above demonstrated the importance of membrane IGF-1R as well as its downstream cascade in retaining/promoting the survival of SKOV3-T, especially when HER2-related signal pathway was down-regulated.Figure 2

Bottom Line: Reversing the resistance often results in better clinical therapeutic effect.It was also confirmed preliminarily that the mechanism of antibody might be to inhibit the activation of IGF-1R and downstream MAPK, AKT pathway transduction.In similar cases, not only acquired but also de novo, good curative effect might be achieved using combined antibody therapy strategies.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Cellular and Molecular Immunology, Institute of Immunology, Henan University, Kaifeng, 475001, China. 1790735517@qq.com.

ABSTRACT

Background: Antibody resistance, not only de novo but also acquired cases, usually exists and is related with lower survival rate and high risk of recurrence. Reversing the resistance often results in better clinical therapeutic effect. Previously, we established a trastuzumab-resistant ovarian cancer cell line, named as SKOV3-T, with lower HER2 and induced higher IGF-1R expression level to keep cell survival.

Methods: IGF-1R was identified important for SKOV3-T growth. Then, a novel anti-IGF-1R monoclonal antibody, named as LMAb1, was used to inhibit SKOV3-T in cell growth/proliferation, migration, clone formation and in vivo carcinogenicity.

Results: In both in vitro and in vivo assays, LMAb1 showed effective anti-tumor function, especially when being used in combination with trastuzumab, which was beneficial to longer survival time of mice as well as smaller tumor. It was also confirmed preliminarily that the mechanism of antibody might be to inhibit the activation of IGF-1R and downstream MAPK, AKT pathway transduction.

Conclusion: We achieved satisfactory anti-tumor activity using trastuzumab plus LMAb1 in trastuzumab-resistant ovarian cancer model. In similar cases, not only acquired but also de novo, good curative effect might be achieved using combined antibody therapy strategies.

Show MeSH
Related in: MedlinePlus