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MGr1-Ag/37LRP induces cell adhesion-mediated drug resistance through FAK/PI3K and MAPK pathway in gastric cancer.

Sun L, Liu L, Liu X, Wang Y, Li M, Yao L, Yang J, Ji G, Guo C, Pan Y, Liang S, Wang B, Ding J, Zhang H, Shi Y - Cancer Sci. (2014)

Bottom Line: However, the MGr1-Ag-initiated intracellular signal transduction pathway is still unknown.Further study found that, as a receptor of ECM components, MGr1-Ag/37LRP may activate the downstream signal pathway PI3K/AKT and MAPK/ERK through interaction with phosphorylated FAK.Moreover, the sensitivity to chemotherapeutic drugs could be significantly enhanced by inhibiting MGr1-Ag/37LRP expression through mAbs, siRNA, and antisense oligonucleotide.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Cancer Biology, Xijing Hospital of Digestive Diseases, Fourth Military Medical University, Xi'an, China.

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Related in: MedlinePlus

(On the previous page) Effect of MGr1-Ag/37LRP mAb, siRNA, and antisense oligonucleotide (ASO) treatment on gastric cancer (GC) drug-resistant cell line SGC7901/VCR tumor growth and chemosensitivity in vitro. (a) SGC7901/VCR cells were treated with indicated concentrations of MGr1-Ag/37LRP ASO or scrambled ASO (SASO) controls, MGr1-Ag/37LRP siRNA or scrambled siRNA (SsiRNA), and mAb MGr1 or control antibody for 48 h. Total protein was extracted from culture cells. (b) Cell adhesion assay. After 2 h of adhesion, the transfected cells and control cells attached to laminin or BSA was counted under a microscope. (c) Sensitivity of GC transfected cells to chemotherapeutic drugs was evaluated using the colony-forming assay. The concentration of each drug that caused a 50% reduction in the number of colonies (IC50) was calculated. (d) Apoptosis indexes of GC cells induced by vincristine (VCR) detected by flow cytometry. *P < 0.05 versus SGC7901/VCR cell incubated with control antibody; #P < 0.05 versus SGC7901/VCR scramble siRNA; ^P <  0.05 versus scramble ASO. Representative experiments are shown from three for each cell type.
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fig04: (On the previous page) Effect of MGr1-Ag/37LRP mAb, siRNA, and antisense oligonucleotide (ASO) treatment on gastric cancer (GC) drug-resistant cell line SGC7901/VCR tumor growth and chemosensitivity in vitro. (a) SGC7901/VCR cells were treated with indicated concentrations of MGr1-Ag/37LRP ASO or scrambled ASO (SASO) controls, MGr1-Ag/37LRP siRNA or scrambled siRNA (SsiRNA), and mAb MGr1 or control antibody for 48 h. Total protein was extracted from culture cells. (b) Cell adhesion assay. After 2 h of adhesion, the transfected cells and control cells attached to laminin or BSA was counted under a microscope. (c) Sensitivity of GC transfected cells to chemotherapeutic drugs was evaluated using the colony-forming assay. The concentration of each drug that caused a 50% reduction in the number of colonies (IC50) was calculated. (d) Apoptosis indexes of GC cells induced by vincristine (VCR) detected by flow cytometry. *P < 0.05 versus SGC7901/VCR cell incubated with control antibody; #P < 0.05 versus SGC7901/VCR scramble siRNA; ^P <  0.05 versus scramble ASO. Representative experiments are shown from three for each cell type.

Mentions: To study the functional role of MGr1-Ag/37LRP in CAM-DR in GC, mAb-, ASO-, or siRNA-induced inhibition of MGr1-Ag/37LRP was determined by Western blot analysis. As shown in Figure4(a), treatment of SGC7901/VCR cells with ASO significantly reduced MGr1-Ag/37LRP protein levels by up to 75% in a dose-dependent manner, whereas MGr1-Ag/37LRP protein expression was not significantly suppressed by scrambled oligonucleotide. Similiar, MGr1-Ag expression was also inhibited at a dose-dependent manner using 10−3–1 nmol/L siRNA and 0.5–10 mg/L mAb. The cell adhesion assay, in vitro drug sensitivity assay, and annexin V/propidium iodide staining were used to exploit the MDR phenotype by blocking MGr1-Ag/37LRP with mAb, siRNA, ASO after adhesion to LN and control component (BSA) (Fig.4b–d). SGC7901/VCR cells transfected with siRNA (1 nmol/L), mAb (10 mg/L), and ASO (40 nM) showed significantly decreased mean adhesion cell number than that of control cells after adhesion to LN. Similarly, SGC7901/VCR cells transfected with siRNA (1 nmol/L), mAb (10 mg/L), and ASO (40 nM) showed significant increased IC50 values of VCR and 5-fluorouracil (5-FU) (Fig.4c), and decreased apoptotic index values in the same conditions (Fig.4d). These results suggested that inhibition of MGr1-Ag expression could partly reverse the CAM-DR phenotype in vitro.


MGr1-Ag/37LRP induces cell adhesion-mediated drug resistance through FAK/PI3K and MAPK pathway in gastric cancer.

Sun L, Liu L, Liu X, Wang Y, Li M, Yao L, Yang J, Ji G, Guo C, Pan Y, Liang S, Wang B, Ding J, Zhang H, Shi Y - Cancer Sci. (2014)

(On the previous page) Effect of MGr1-Ag/37LRP mAb, siRNA, and antisense oligonucleotide (ASO) treatment on gastric cancer (GC) drug-resistant cell line SGC7901/VCR tumor growth and chemosensitivity in vitro. (a) SGC7901/VCR cells were treated with indicated concentrations of MGr1-Ag/37LRP ASO or scrambled ASO (SASO) controls, MGr1-Ag/37LRP siRNA or scrambled siRNA (SsiRNA), and mAb MGr1 or control antibody for 48 h. Total protein was extracted from culture cells. (b) Cell adhesion assay. After 2 h of adhesion, the transfected cells and control cells attached to laminin or BSA was counted under a microscope. (c) Sensitivity of GC transfected cells to chemotherapeutic drugs was evaluated using the colony-forming assay. The concentration of each drug that caused a 50% reduction in the number of colonies (IC50) was calculated. (d) Apoptosis indexes of GC cells induced by vincristine (VCR) detected by flow cytometry. *P < 0.05 versus SGC7901/VCR cell incubated with control antibody; #P < 0.05 versus SGC7901/VCR scramble siRNA; ^P <  0.05 versus scramble ASO. Representative experiments are shown from three for each cell type.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4317895&req=5

fig04: (On the previous page) Effect of MGr1-Ag/37LRP mAb, siRNA, and antisense oligonucleotide (ASO) treatment on gastric cancer (GC) drug-resistant cell line SGC7901/VCR tumor growth and chemosensitivity in vitro. (a) SGC7901/VCR cells were treated with indicated concentrations of MGr1-Ag/37LRP ASO or scrambled ASO (SASO) controls, MGr1-Ag/37LRP siRNA or scrambled siRNA (SsiRNA), and mAb MGr1 or control antibody for 48 h. Total protein was extracted from culture cells. (b) Cell adhesion assay. After 2 h of adhesion, the transfected cells and control cells attached to laminin or BSA was counted under a microscope. (c) Sensitivity of GC transfected cells to chemotherapeutic drugs was evaluated using the colony-forming assay. The concentration of each drug that caused a 50% reduction in the number of colonies (IC50) was calculated. (d) Apoptosis indexes of GC cells induced by vincristine (VCR) detected by flow cytometry. *P < 0.05 versus SGC7901/VCR cell incubated with control antibody; #P < 0.05 versus SGC7901/VCR scramble siRNA; ^P <  0.05 versus scramble ASO. Representative experiments are shown from three for each cell type.
Mentions: To study the functional role of MGr1-Ag/37LRP in CAM-DR in GC, mAb-, ASO-, or siRNA-induced inhibition of MGr1-Ag/37LRP was determined by Western blot analysis. As shown in Figure4(a), treatment of SGC7901/VCR cells with ASO significantly reduced MGr1-Ag/37LRP protein levels by up to 75% in a dose-dependent manner, whereas MGr1-Ag/37LRP protein expression was not significantly suppressed by scrambled oligonucleotide. Similiar, MGr1-Ag expression was also inhibited at a dose-dependent manner using 10−3–1 nmol/L siRNA and 0.5–10 mg/L mAb. The cell adhesion assay, in vitro drug sensitivity assay, and annexin V/propidium iodide staining were used to exploit the MDR phenotype by blocking MGr1-Ag/37LRP with mAb, siRNA, ASO after adhesion to LN and control component (BSA) (Fig.4b–d). SGC7901/VCR cells transfected with siRNA (1 nmol/L), mAb (10 mg/L), and ASO (40 nM) showed significantly decreased mean adhesion cell number than that of control cells after adhesion to LN. Similarly, SGC7901/VCR cells transfected with siRNA (1 nmol/L), mAb (10 mg/L), and ASO (40 nM) showed significant increased IC50 values of VCR and 5-fluorouracil (5-FU) (Fig.4c), and decreased apoptotic index values in the same conditions (Fig.4d). These results suggested that inhibition of MGr1-Ag expression could partly reverse the CAM-DR phenotype in vitro.

Bottom Line: However, the MGr1-Ag-initiated intracellular signal transduction pathway is still unknown.Further study found that, as a receptor of ECM components, MGr1-Ag/37LRP may activate the downstream signal pathway PI3K/AKT and MAPK/ERK through interaction with phosphorylated FAK.Moreover, the sensitivity to chemotherapeutic drugs could be significantly enhanced by inhibiting MGr1-Ag/37LRP expression through mAbs, siRNA, and antisense oligonucleotide.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Cancer Biology, Xijing Hospital of Digestive Diseases, Fourth Military Medical University, Xi'an, China.

Show MeSH
Related in: MedlinePlus