Effects Of Lamprey PHB2 On Cell Proliferation And Migration In Non-small Cell Lung Cancer

According to the world cancer report issued by the World Health Organization(WHO),lung cancer accounts for about 20% of total cancer deaths.Among them,80% of lung cancer belongs to non-small cell lung cancer(NSCLC),30% of which has mutations in the proto-oncogene KRAS.There are a lot of NSCLC patients with KRAS mutation,but there are few effective clinical treatments.Therefore,it is particularly important to carry out the study on NSCLC with KRAS mutation.There are two homologous subtypes of Prohibitin in eukaryotes,namely Prohibitin 1(PHB1)and Prohibitin 2(PHB2).In recent years,studies have shown that PHB2 is closely related to the occurrence and development of tumors,so the value of PHB2 in cancer treatment deserves further study.Lamprey is an important model organism for studying vertebrate evolution and adaptive immunity.In contrast to the extensive study of mammal PHB2,there are few studies on lamprey PHB2.In this study,the expression of PHBs in NCI-H1650 cells,A549 cells,Calu-1 cells and NCI-H226 cells of NSCLC was firstly explored by q PCR,Western blot and cell immunofluorescence.It was found that PHBs was highly expressed in NCI-H1650 cells,A549 cells and Calu-1 cells.Based on the previous work of our research group,the purified r Lm-PHB2(Recombinant lamprey PHB2 protein)was incubated with the above four cells,and cell proliferation and cell cycle were detected by CCK method and flow cytometry,respectively.The results showed that r Lm-PHB2 blocked the cells in the G2 phase and inhibited the proliferation of A549 cells,Calu-1 cells and NCI-H226 cells with KRAS mutation,among which the effect on Calu-1 cells was the most obvious,showing a concentration and time dependence.The eukaryotic over-expression vector p EGFP-N1-LmPHB2 was transfected into four kinds of cells to detect the effect of Lm-PHB2 on cell proliferation and cell cycle.The results showed that the endogenous expression of Lm-PHB2 blocked the cells in the G2 phase and inhibited the proliferation of A549 cells and Calu-1cells.The results of q PCR and Western blot showed that Lm-PHB2 not only down-regulated the expression of CDC25 C,CCNB1,PLK1 and Wee1,but also reduced the phosphorylation level of CCNB1 and CDC25 C,which blocked Calu-1 cells in G2 / M phase.Subsequently,the effect of Lm-PHB2 on cell migration was detected by scratch test,and it was found that Lm-PHB2 could inhibit the migration rate of Calu-1 cells.Western blot was used to detect theexpression of E-Cadherin and Vimentin,and it was found that Lm-PHB2 could inhibit the epithelial mesenchymal transition of Calu-1 cells,and then inhibit their migration.In this study,it was found that Lm-PHB2 could not only inhibit the proliferation of NSCLC A549 cells and Calu-1 cells by inhibiting the expression and phosphorylation levels of cycle-related proteins,but also inhibit the migration rate of Calu-1 cells.However,the molecular mechanism of the action remains to be further studied.This study provides clues for the early diagnosis and targeted drug development of NSCLC.