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Homoharringtonine induces apoptosis and inhibits STAT3 via IL-6/JAK1/STAT3 signal pathway in Gefitinib-resistant lung cancer cells.

Cao W, Liu Y, Zhang R, Zhang B, Wang T, Zhu X, Mei L, Chen H, Zhang H, Ming P, Huang L - Sci Rep (2015)

Bottom Line: Unfortunately, treatment with Gefitinib for a period of time will result in drug resistance and cause treatment failure in clinic.NCI-H1975 cells with EGFR T790M mutation are more sensitive to HHT treatment compared with that of A549 cells with wild type EGFR.HHT reversiblely inhibited IL-6-induced STAT3 Tyrosine 705 phosphorylation and reduced anti-apoptotic proteins expression.

View Article: PubMed Central - PubMed

Affiliation: 1] School of Life Sciences, Tsinghua University, Beijing, 100084, China [2] The Shenzhen Key Laboratory of Gene and Antibody Therapy, State Key Laboratory of Health Science and Technology (prep), Center for Biotechnology &Biomedicine and Division of Life &Health Sciences, Graduate School at Shenzhen, Tsinghua University, Shenzhen, Guangdong, 518055, China.

ABSTRACT
Tyrosine kinase inhibitors (TKIs) are mostly used in non-small cell lung cancer (NSCLC) treatment. Unfortunately, treatment with Gefitinib for a period of time will result in drug resistance and cause treatment failure in clinic. Therefore, exploring novel compounds to overcome this resistance is urgently required. Here we investigated the antitumor effect of homoharringtonine (HHT), a natural compound extracted from Cephalotaxus harringtonia, on Gefitinib-resistant NSCLC cell lines in vitro and in vivo. NCI-H1975 cells with EGFR T790M mutation are more sensitive to HHT treatment compared with that of A549 cells with wild type EGFR. HHT inhibited cells growth, cell viability and colony formation, as well as induced cell apoptosis through mitochondria pathway. Furthermore, we explored the mechanism of HHT inhibition on NSCLC cells. Higher level of interleukin-6 (IL-6) existed in lung cancer patients and mutant EGFR and TGFβ signal requires the upregulation of IL-6 through the gp130/JAK pathway to overactive STAT3, an oncogenic protein which has been considered as a potential target for cancer therapy. HHT reversiblely inhibited IL-6-induced STAT3 Tyrosine 705 phosphorylation and reduced anti-apoptotic proteins expression. Gefitinib-resistant NSCLC xenograft tests also confirmed the antitumor effect of HHT in vivo. Consequently, HHT has the potential in Gefitinib-resistant NSCLC treatment.

No MeSH data available.


Related in: MedlinePlus

HHT exerts synergistic effect combining with docetaxel.(A and B): A549 (A) and H1975 (B) cells were treated for 24 h with DTX and/or HHT, and then assessed by MTT assay. (C): A549 and H1975 cells were treated with HHT (2 μM or 0.25 μM), Gefitinib (2 mM), DTX (8 nM) alone or together for 24 h. The treated cells were collected, lysed and assessed by western blot with indicated antibodies. The blots shown are derived from multiple gels. Membrane was cut based on the molecular weight, probed with antibody of interest and band of interest is indicated with an arrow. All the full-length blots are presented in Supplementary Figure 4.
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f6: HHT exerts synergistic effect combining with docetaxel.(A and B): A549 (A) and H1975 (B) cells were treated for 24 h with DTX and/or HHT, and then assessed by MTT assay. (C): A549 and H1975 cells were treated with HHT (2 μM or 0.25 μM), Gefitinib (2 mM), DTX (8 nM) alone or together for 24 h. The treated cells were collected, lysed and assessed by western blot with indicated antibodies. The blots shown are derived from multiple gels. Membrane was cut based on the molecular weight, probed with antibody of interest and band of interest is indicated with an arrow. All the full-length blots are presented in Supplementary Figure 4.

Mentions: In order to improve therapeutic efficacy and reduce the drug toxicity to patients or/and non-target tissues, one or more other drugs usually are used in clinic. Furthermore, an optimal combined treatment may decrease or postpone the drug resistance45. Docetaxel (DTX) can bind to microtubules, cause cell-cycle arrest and apoptosis and therefore is approved to cure a certain of tumors including NSCLC46. We examined whether HHT and DTX combination had the synergistic effect in NSCLC treatment. By MTT assay, HHT-induced cell proliferation inhibition was significantly elevated by DTX at 2 nM to 8 nM (Fig. 6A,B). We also analyzed the CI value by the formula47 assisted with CalcuSyn software (Version 2.1) and found that the CI values were less than 1 (Table 1), which indicated that HHT and DTX played synergistic effect in Gefitinib-resistant NSCLC cells. By western blot, we further confirmed the synergistic effect and found that HHT and DTX combination resulted in elevated levels of Caspase 3 activation, cleavage of PARP and decreased level of STAT3 phosphorylation (Fig. 6C).


Homoharringtonine induces apoptosis and inhibits STAT3 via IL-6/JAK1/STAT3 signal pathway in Gefitinib-resistant lung cancer cells.

Cao W, Liu Y, Zhang R, Zhang B, Wang T, Zhu X, Mei L, Chen H, Zhang H, Ming P, Huang L - Sci Rep (2015)

HHT exerts synergistic effect combining with docetaxel.(A and B): A549 (A) and H1975 (B) cells were treated for 24 h with DTX and/or HHT, and then assessed by MTT assay. (C): A549 and H1975 cells were treated with HHT (2 μM or 0.25 μM), Gefitinib (2 mM), DTX (8 nM) alone or together for 24 h. The treated cells were collected, lysed and assessed by western blot with indicated antibodies. The blots shown are derived from multiple gels. Membrane was cut based on the molecular weight, probed with antibody of interest and band of interest is indicated with an arrow. All the full-length blots are presented in Supplementary Figure 4.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4499885&req=5

f6: HHT exerts synergistic effect combining with docetaxel.(A and B): A549 (A) and H1975 (B) cells were treated for 24 h with DTX and/or HHT, and then assessed by MTT assay. (C): A549 and H1975 cells were treated with HHT (2 μM or 0.25 μM), Gefitinib (2 mM), DTX (8 nM) alone or together for 24 h. The treated cells were collected, lysed and assessed by western blot with indicated antibodies. The blots shown are derived from multiple gels. Membrane was cut based on the molecular weight, probed with antibody of interest and band of interest is indicated with an arrow. All the full-length blots are presented in Supplementary Figure 4.
Mentions: In order to improve therapeutic efficacy and reduce the drug toxicity to patients or/and non-target tissues, one or more other drugs usually are used in clinic. Furthermore, an optimal combined treatment may decrease or postpone the drug resistance45. Docetaxel (DTX) can bind to microtubules, cause cell-cycle arrest and apoptosis and therefore is approved to cure a certain of tumors including NSCLC46. We examined whether HHT and DTX combination had the synergistic effect in NSCLC treatment. By MTT assay, HHT-induced cell proliferation inhibition was significantly elevated by DTX at 2 nM to 8 nM (Fig. 6A,B). We also analyzed the CI value by the formula47 assisted with CalcuSyn software (Version 2.1) and found that the CI values were less than 1 (Table 1), which indicated that HHT and DTX played synergistic effect in Gefitinib-resistant NSCLC cells. By western blot, we further confirmed the synergistic effect and found that HHT and DTX combination resulted in elevated levels of Caspase 3 activation, cleavage of PARP and decreased level of STAT3 phosphorylation (Fig. 6C).

Bottom Line: Unfortunately, treatment with Gefitinib for a period of time will result in drug resistance and cause treatment failure in clinic.NCI-H1975 cells with EGFR T790M mutation are more sensitive to HHT treatment compared with that of A549 cells with wild type EGFR.HHT reversiblely inhibited IL-6-induced STAT3 Tyrosine 705 phosphorylation and reduced anti-apoptotic proteins expression.

View Article: PubMed Central - PubMed

Affiliation: 1] School of Life Sciences, Tsinghua University, Beijing, 100084, China [2] The Shenzhen Key Laboratory of Gene and Antibody Therapy, State Key Laboratory of Health Science and Technology (prep), Center for Biotechnology &Biomedicine and Division of Life &Health Sciences, Graduate School at Shenzhen, Tsinghua University, Shenzhen, Guangdong, 518055, China.

ABSTRACT
Tyrosine kinase inhibitors (TKIs) are mostly used in non-small cell lung cancer (NSCLC) treatment. Unfortunately, treatment with Gefitinib for a period of time will result in drug resistance and cause treatment failure in clinic. Therefore, exploring novel compounds to overcome this resistance is urgently required. Here we investigated the antitumor effect of homoharringtonine (HHT), a natural compound extracted from Cephalotaxus harringtonia, on Gefitinib-resistant NSCLC cell lines in vitro and in vivo. NCI-H1975 cells with EGFR T790M mutation are more sensitive to HHT treatment compared with that of A549 cells with wild type EGFR. HHT inhibited cells growth, cell viability and colony formation, as well as induced cell apoptosis through mitochondria pathway. Furthermore, we explored the mechanism of HHT inhibition on NSCLC cells. Higher level of interleukin-6 (IL-6) existed in lung cancer patients and mutant EGFR and TGFβ signal requires the upregulation of IL-6 through the gp130/JAK pathway to overactive STAT3, an oncogenic protein which has been considered as a potential target for cancer therapy. HHT reversiblely inhibited IL-6-induced STAT3 Tyrosine 705 phosphorylation and reduced anti-apoptotic proteins expression. Gefitinib-resistant NSCLC xenograft tests also confirmed the antitumor effect of HHT in vivo. Consequently, HHT has the potential in Gefitinib-resistant NSCLC treatment.

No MeSH data available.


Related in: MedlinePlus