OBJECTIVE To investigate 3-O-(Z)-coumaroyloleanolic acid(3-COA),an active ingredient of oleanolic acid in the TAIWANG leaves of E.oldhamii Maxim,that has been shown to have antitumor activity against A549 lung cancer ...OBJECTIVE To investigate 3-O-(Z)-coumaroyloleanolic acid(3-COA),an active ingredient of oleanolic acid in the TAIWANG leaves of E.oldhamii Maxim,that has been shown to have antitumor activity against A549 lung cancer cells,overcomes Cks1b-induced chemoresistance in lung cancer by inhibiting Hsp90 and MEK pathways.METHODS Cisplatin(CDDP)and doxorubicin(DOX)sensitivity was assessed through proliferation,viability,and clonogenic potential induction in cells overexpressing Cks1b(Cks1b-OE).The mechanism for resistance and 3-COA sensitivity were elucidated by immunoblot analysis of Hsp90 and MEK,and confirmed by sh RNA knockdown.Inhibition of 3-COA or3-COA combined with CDDP(3-COA&CDDP)was assayed in early primary lung cancer(IPH),late primary lung cancer(RFH)cells,and tumor-burdened immunodeficiency mice in vivo.RESULTS The ectopic overexpression of Cks1b in human lung cancer cells induces chemoresistance of the cells to CDDP and DOX,but not3-COA,through mechanisms independent of its canonical Skp2-p27 pathway.Further dissection with application of shR NA and selective inhibitors reveals that Hsp90 and MEK1/2 are the critical components of the non-canonical pathways responsible for the Cks1b-induced chemoresistance.Interestingly,inhibition of either Hsp90 or MEK1/2rendered a similar magnitude of antitumor activity by resensitization of the chemoresistant Cks1b-OE cells to CDDP and DOX,suggesting that both Hsp90 and MEK1/2are essentialto Cks1b for induction of chemoresistance.IC50of 3-COA is 6.82μmol·L-1in H358 Cks1b and8.22μmol·L-1in H226 Cks1b,which were not significantly higher than those in H358 EV and H226 EV,respectively.Furthermore,3-COA mimicked PU-H71,a Hsp90-specific inhibitor,targeted Hsp90 and MEK to reduce the expression of their downstream,respectively.Importantly,compared with CDDP treatment,3-COA or 3-COA&CDDP signifi-cantly inhibited RFH cells in vitro.Moreover,3-COA or3-COA&CDDP significantly prolonged more survival for a H358 Cks1b-OE inducing tumor-burdened mice than PD98059(a MEK-specific inhibitor)in vivo.CONCLUSION Our data report for the first time that Cks1b employs Hsp90 and MEK1/2 pathways in lung cancer cells to develop chemoresistance and identify 3-COA as a potential antitumor drug for clinical treatment of chemoresistant lung cancer.展开更多
Objective The aim of this study was to explore the clinical significance of the expression of proteins human bone marrow endothelial cell markers(HBME-1), Galectin-3, and cytokeratin19(CK19), as well as the status of ...Objective The aim of this study was to explore the clinical significance of the expression of proteins human bone marrow endothelial cell markers(HBME-1), Galectin-3, and cytokeratin19(CK19), as well as the status of v-raf murine sarcoma viral oncogene homolog B1(BRAF) mutation in papillary thyroid carcinoma(PTC). Methods Immunohistochemical staining was performed in 82 specimens each of PTC and papillary benign lesions to detect the expression of HBME-1, Galectin-3, and CK19. Polymerase chain reaction(PCR) and gene sequencing were performed on 60 specimens each of PTC and papillary benign lesions to detect the status of BRAF mutation. Results The positive expression ratios of HBME-1, Galectin-3, and CK19 in PTC were 98.8%, 97.6% and 100% respectively, which were significantly higher than the expressions in papillary benign lesions(P < 0.05). No significant relationship was observed between the expression of these makers and the clinicopathological features of PTC. The sensitivity of co-expression of HBME-1 and CK19 or HBME-1 and Galectin-3 as diagnostic criteria of PTC was 99.9%, with a specificity of 95.4%. BRAF mutation was detected in 40 of 60 PTC(66.7%) specimens. There was a statistical difference in BRAF mutations between PTC and papillary benign lesions(P < 0.05); there were no associations between BRAF mutation and the clinicopathological features of PTC. Conclusion Combined immunohistochemical staining of HBME-1, Galectin-3, and CK19 can further improve the sensitivity and specificity of differential diagnosis of PTC. BRAF mutation is a significant genetic event, which may have diagnostic value for PTC.展开更多
基金The project supported by National Natural Science Foundation of China(81402023)Natural Science Foundation of Guangdong Province(2014A030310022)+1 种基金Scientific Research Projects of Guangzhou(201510010212)College Students Cultivate Special Science and Technology Innovation Projects of Guangdong in 2016(pdjh2016b0408)
文摘OBJECTIVE To investigate 3-O-(Z)-coumaroyloleanolic acid(3-COA),an active ingredient of oleanolic acid in the TAIWANG leaves of E.oldhamii Maxim,that has been shown to have antitumor activity against A549 lung cancer cells,overcomes Cks1b-induced chemoresistance in lung cancer by inhibiting Hsp90 and MEK pathways.METHODS Cisplatin(CDDP)and doxorubicin(DOX)sensitivity was assessed through proliferation,viability,and clonogenic potential induction in cells overexpressing Cks1b(Cks1b-OE).The mechanism for resistance and 3-COA sensitivity were elucidated by immunoblot analysis of Hsp90 and MEK,and confirmed by sh RNA knockdown.Inhibition of 3-COA or3-COA combined with CDDP(3-COA&CDDP)was assayed in early primary lung cancer(IPH),late primary lung cancer(RFH)cells,and tumor-burdened immunodeficiency mice in vivo.RESULTS The ectopic overexpression of Cks1b in human lung cancer cells induces chemoresistance of the cells to CDDP and DOX,but not3-COA,through mechanisms independent of its canonical Skp2-p27 pathway.Further dissection with application of shR NA and selective inhibitors reveals that Hsp90 and MEK1/2 are the critical components of the non-canonical pathways responsible for the Cks1b-induced chemoresistance.Interestingly,inhibition of either Hsp90 or MEK1/2rendered a similar magnitude of antitumor activity by resensitization of the chemoresistant Cks1b-OE cells to CDDP and DOX,suggesting that both Hsp90 and MEK1/2are essentialto Cks1b for induction of chemoresistance.IC50of 3-COA is 6.82μmol·L-1in H358 Cks1b and8.22μmol·L-1in H226 Cks1b,which were not significantly higher than those in H358 EV and H226 EV,respectively.Furthermore,3-COA mimicked PU-H71,a Hsp90-specific inhibitor,targeted Hsp90 and MEK to reduce the expression of their downstream,respectively.Importantly,compared with CDDP treatment,3-COA or 3-COA&CDDP signifi-cantly inhibited RFH cells in vitro.Moreover,3-COA or3-COA&CDDP significantly prolonged more survival for a H358 Cks1b-OE inducing tumor-burdened mice than PD98059(a MEK-specific inhibitor)in vivo.CONCLUSION Our data report for the first time that Cks1b employs Hsp90 and MEK1/2 pathways in lung cancer cells to develop chemoresistance and identify 3-COA as a potential antitumor drug for clinical treatment of chemoresistant lung cancer.
基金This work was supported by National Institutes of Health grants CA55819 (JDS, FZ, and BB) and CA97513 (JDS) and by the Fund to Cure Myeloma and Peninsula Community Foundation
文摘Objective The aim of this study was to explore the clinical significance of the expression of proteins human bone marrow endothelial cell markers(HBME-1), Galectin-3, and cytokeratin19(CK19), as well as the status of v-raf murine sarcoma viral oncogene homolog B1(BRAF) mutation in papillary thyroid carcinoma(PTC). Methods Immunohistochemical staining was performed in 82 specimens each of PTC and papillary benign lesions to detect the expression of HBME-1, Galectin-3, and CK19. Polymerase chain reaction(PCR) and gene sequencing were performed on 60 specimens each of PTC and papillary benign lesions to detect the status of BRAF mutation. Results The positive expression ratios of HBME-1, Galectin-3, and CK19 in PTC were 98.8%, 97.6% and 100% respectively, which were significantly higher than the expressions in papillary benign lesions(P < 0.05). No significant relationship was observed between the expression of these makers and the clinicopathological features of PTC. The sensitivity of co-expression of HBME-1 and CK19 or HBME-1 and Galectin-3 as diagnostic criteria of PTC was 99.9%, with a specificity of 95.4%. BRAF mutation was detected in 40 of 60 PTC(66.7%) specimens. There was a statistical difference in BRAF mutations between PTC and papillary benign lesions(P < 0.05); there were no associations between BRAF mutation and the clinicopathological features of PTC. Conclusion Combined immunohistochemical staining of HBME-1, Galectin-3, and CK19 can further improve the sensitivity and specificity of differential diagnosis of PTC. BRAF mutation is a significant genetic event, which may have diagnostic value for PTC.
