ABSTRACT
45.7% of Chinese patients with advanced lung adenocarcinoma were reported to harbour sensitizing epidermal growth factor receptor (EGFR) mutations. Limited therapeutic options are left for non-small cell lung cancer (NSCLC) harbouring sensitizing EGFR mutations after failure of EGFR-tyrosine kinase inhibitor (TKI) therapy and chemotherapy, finding effective options for them is an unmet clinic need. Herein we reported a case that till January 12, 2021, an 82-year-old female with sensitizing EGFR-mutant advanced lung adenocarcinoma received a surprising progression-free survival (PFS) benefit of over 21 months from the combination therapy of pembrolizumab and anlotinib after her failure of treatments of osimertinib, chemotherapy and anlotinib-monotherapy. .
Subject(s)
Aged, 80 and over , Female , Humans , Adenocarcinoma of Lung/genetics , Antibodies, Monoclonal, Humanized , Antineoplastic Combined Chemotherapy Protocols , Carcinoma, Non-Small-Cell Lung/genetics , ErbB Receptors/genetics , Indoles , Lung Neoplasms/genetics , Mutation , QuinolinesABSTRACT
Objective: To investigate the expression and expression efficiency of recombinant adeno-associated viral containing human thyrotropin receptor A subunit (rAAV2/9-hTSHR289-IRES-ZS Green) in mouse embryonic fibroblasts (NIH3T3) or in mice. Methods: NIH3T3 was infected by rAAV2/9-hTSHR289-IRES-ZsGreen and its TSHR289 protein expression was detected by Western blotting. Female BALB/c mice were intramuscularly injected with different doses of rAAV2/9-hTSHR289-IRES-ZsGreen; the expressions of the target protein in different organs were determined by immunofluorescence while the serum TSHR antibody (TRAb) titer was determined by radioimmunoassay. Results: NIH3T3 cells transfected with rAAV2/9-hTSHR289-IRES-ZsGreen expressed hTSHR289 protein both at 48 h and 72 h. The expression of target protein at 48 h or 72 h was significantly higher than that at 24 h (P<0.05). There were different levels of TSHR289 expression in leg skeletal muscle, heart, liver and spleen under fluorescence microscope. The results of radioimmunoassay showed that the higher dose injection produced a higher titer of TSHR antibody, but only the TRAb level in high dose and control injection exhibited a statistical difference (P<0.05) at week 4, week 8 and week 12. Furthermore, strong antibody response was observed in the mice injected with high dose and medium dose of rAAV2/9-hTSHR289-IRES-ZsGreen at week 4 and gradually weakened between 4 and 8 weeks. In addition, the antibody lasted for 12 weeks. Results: rAAV2/9-hTSHR289-IRES-ZsGreen can highly express hTSHR289 protein in vivo and in vitro, and the hTSHR289 protein displays strong immunogenicity. It indicates that rAAV2/9-hTSHR289-IRES-ZsGreen might become a more effective means of preparing Graves' disease model.
ABSTRACT
Objective To explore the effect of Yes-associated protein 1 (YAP1) and potential mechanism on erlotinib ( ER) resistance in lung adenocarcinoma.Methods In PC-9 cells and acquired ER resistant PC-9 ( PC-9/ER) cells, the expression changes in YAP1 gene were measured by quantitative real-time PCR( RT-qPCR) and Western blot.Indirect immunofluorescence was adopted to observe the location of YAP1.PC-9/ER cells were treated with the verteporfin ( VP, YAP1 inhibitor) for 24 h and 48 h, respectively.Expression changes in mRNA and proteins of YAP1, AKT and p-AKT were detected in the presence or absence of VP.The effect of VP was analyzed by drug resistance index using Cell Counting Kit 8(CCK-8) assay.Results The resistance index of PC-9/ER cells was (99.80 ±25.81).Compared with PC-9 cells, the expression levels of YAP1 mRNA and protein were increased in PC-9/ER.The inhibitory efficiency of VP was (50.96 ±5.86)%, and the levels of AKT and p-AKT proteins were down-regulated by the inhibition of YAP1 simultaneously.The half maximal inhibitory concentration (IC50) of PC-9/ER decreased from (11.10 ±2.72) to (1.47 ± 0.32)μmol/L (P =0.024).Resistance index was reduced to one eighth of the original.Conclusion These results indicate that the YAP1 mediates ER resistance in lung adenocarcinoma.Suppression of YAP1 can reduce the resistance through PI3K/AKT signaling pathway.Therefore, YAP1 may be a potential target for lung cancer gene therapy.