A Study On The Effect Of Anlotinib On Lung Carcinoma Cell Line After EGFR-TKI Resistance

ObjectiveThe incidence and fatality rate of Non-small cell lung carcinoma(NSCLC)remained high all over the world.At present,the combined treatment of chemotherapeutic agents cannot significantly prolong the survival period of patients.Although the generation of targeted therapy has improved the quality of life of patients with advanced NSCLC,the challenge of acquired resistance would inevitably appear that lead to the progression of disease and poor outcome.According to the unsatisfied result,it is necessary for researchers to explore more effective therapeutic drugs to improve the prognosis of patients with advanced NSCLC.Anlotinib is a novel,oral multitarget tyrosine kinase inhibitor for tumor angiogenesis and proliferative signaling.The data from ALTER 0303 trial showed that considerable improvement in overall survival and progression-free survival was observed in the anlotinib group over the placebo group as a third line treatment for Non-small cell lung cancer.However,there was a lack of data on the treatment of patients with advanced NSCLC who undergone the resistance of first generation EGFR TKI.Whether it can bring more options of second-line therapy for patients remains to be further confirmed.What’s more,the combined treatment of Anlotinib and chemotherapy agents was rare reported in clinic.Whether combination therapy could generate synergistic effect or which chemotherapy drug was compatible with Anlotinib were problems we need to consider and solve.The purpose of our study was to confirm the efficacy of anlotinib as a second-line treatment in patients with advanced NSCLC after first-line epidermal growth factor receptor tyrosine kinase inhibitor resistance by investigating the inhibitory effect of Anlotinib on icotinib resistant NSCLC cell lines and preliminarily evaluated the feasibility of Anlotinib plus chemotherapy agents through the comparison of the proliferation inhibition of different chemotherapy drugs combined with Anlotinib.Methods1.The influence of Anlotinib to the activity of HCC827/IR was detected by MTT assay.2.The HCC827/IR cell lines were exposed to Anlotinib at various concentrations and then stained with DAPI.The changes of morphology of cells were observed by fluorescence microscopy.3.The expression of apoptosis related protein named caspase-3 which generated from HCC827/IR cells after treated with Anlotinib was evaluated by Western blot.4.We compared the proliferation inhibitory effect of Anlotinib combined different chemotherapy drugs on HCC827/IR cells.Results1.Anlotinib could significantly inhibit the proliferation of HCC827/IR cell lines.The IC50 value of anlotinib was 1.66 u M.2.A typical apoptotic morphological changes was observed in nucleus of HCC827/IR cell lines after exposed to Anlotinib 48 hours.3.Anlotinib could induce the apoptosis of HCC827/IR cells through enhancing the expression of caspase-3.While the concentration of Anlotinib increasing,the expression level of caspase-3 was also upregulating.4.It was indicated from the cell inhibition rate curve that Anlotinib combined with docetaxel could inhibit the proliferation of HCC827/IR cells more effectively than Anlotinib plus cisplatin.ConclusionAnlotinib could inhibit the proliferation of HCC827/IR cell lines and induce the apoptosis,which was hopeful to become the second-line therapy for patients with advanced NSCLC who have undergone the failure of first-line EGFR TKI treatment.It was feasible to choose docetaxel as a combined drugs with Anlotinib.