Study Of Pan-HER Inhibitoralone Or Combination On Resistance Non-small Cell Lung Cancer H1975Cell In Vitro

Objective: To study the effect of afatinib, a Pan-HER inhibitor, aloneor combination with PF-04691502, a phosphoinositide-3kinase(PI3K)inhibitor，andtrematinib (GSK1120212), amitogen-activated protein kinasekinase(MEK) inhibitor on Non-Small Cell Lung Cancer(NSCLC) cell lineH1975in vitro.Methods:(1)Compare the proliferation inhibiton effect of gefitinib andlapatinib, EGFR reversible inhibitor, and afatinib, Pan-HER irreversibleinhibitor respectively on resistance NSCLC cell line H1975and EGFRexpressive gastric carcinoma cell line N87by MTS.(2)To analyze PF and afatinib alone or combination effect ofproliferation and migration on resistance NSCLC cell line H1975by MTS，cell scratch healing experiment, flow cytometry measure cell cycle and cellapoptosis.(3)To analyze trematinib and afatinib alone or combination effect ofproliferation and migration on resistance NSCLC cell line H1975by MTS,flow cytometry measure cell cycle and cell apoptosis, cell reactive oxygen (ROS) concertration detect.Results:(1)The proliferation inhibiton effect of gefitinib, lapatinib andafatinib for N87cell line are stronger than the H1975cell line, even theproliferation inhibiton effect of afatinib is stronger than gefitinib andlapatinib in the two cell lines.(2)PF alone inhibit the proliferation of H1975cell line, inhibition tostrengthen with the increase of concentration. Significant synergisticinhibition effects were observed in H1975cell proliferation and migrationwhen afatinib combination with PF. The alone effect of afatinib and PFprimary induce cell G1arrest(P<0.05), and the combination effect inducecell G1and G2arrest(P<0.05). Compared with control group, the apoptosisrate was increased in single drug dosage group of afatinib and PF, evenapoptosis rate was remarkable increased in combination group(P<0.05).(3)Significant synergistic inhibition effects were observed incombination group of afatinib and trematinib. Afatinib and trematinibinduce cell G1arrest(P<0.05). compared with control group and singledrug dosage group of afatinib and trematinib, the apoptosis rate ofcombination group were remarkable increased(P<0.05). And the ROSconcentration of combination group also remarkable increased, whencompared with control group and single drug dosage group of afatinib andtrematinib(P<0.05).Conclusions:(1) The proliferation inhibiton effect of EGFR reversible inhibitor and Pan-HER irreversible inhibitor for N87cell line, which isgastric carcinoma with EGFR high express, are stronger than the resistanceNSCLC cell line H1975, even the proliferation inhibiton effect of Pan-HERinhibitor shows obvious advantage for two cell lines, compared with EGFRreversible inhibitor.(2) Afatinib, a Pan-HER inhibitor, combination with PF, a PI3Kinhibitor, shows synergistic inhibition effects of proliferation, migration andapoptosis on resistance NSCLC cell line H1975, it may associate with cellcycle G1arrest.(3) Afatinib, a Pan-HER inhibitor, combination with trematinib, aMEK inhibitor, shows synergistic inhibition effects of proliferation andapoptosis on resistance NSCLC cell line H1975, it may associate with cellcycle G1arrest and increase the cell ROS concentration etc.(4) Pan-HER inhibitor in combination with PI3K inhibitor and MEKinhibitor can increase the antitumor activity of resistance NSCLC cell lineH1975in vitro, it shows certain guiding significance to the clinic drugcombination.