Research On The Correlation Between PHB-1 And The Biological Activity Of Lung Squamous Cell Carcinoma

Objective:Prohibitins(PHBs),including Prohibitin-1(PHB-1)and Prohibitin-2(PHB-2).PHBs are relatively conservative in structure and are found in the nucleus,Cell membrane,cytoplasm and mitochondria and other subcellular structures are commonly expressed.The function of PHBs is related to its subcellular localization,It involved in cell cycle,mitochondrial structure and function,gene transcription and signal transduction regulation.PHBS are involved in the regulation of many signaling pathways,such as p53,Akt,RAF,Rb,E2F,etc.,so they are very important in the development of various diseases,especially in tumors.Lung cancer is the tumor that has the highest morbidity and mortality.It is divided into two types:small cell lung cancer and non-small cell lung cancer.The most common form of non-small cell lung cancer is squamous cell carcinoma.Up to now,it has been found that PHBs are abControlly expressed in gastric cancer,prostate cancer,breast cancer,esophageal squamous cell carcinoma,colorectal cancer,bladder cancer and other tumors.Early study in our laboratory have found that the expression of PHB-1 in lung squamous cell carcinoma tissues was significantly increased.Therefore,the main purpose of this study is to further explore the effect of PHB-1 on the growth,proliferation,migration and invasion of lung squamous cell carcinoma cells.This will provide new ideas and new methods for the early diagnosis and target therapy of lung squamous cell carcinoma.Methods:SK-MES-1 cells(lung squamous cell carcinoma cell line)were transfected with plasmid to construct stable SK-MES-1 cell lines with lower or higher expression of PHBs.The expression of PHB-1 in three SK-MES-1 cell lines was verified by Real-Time Quantitative PCR.The effects of PHB-1 on cell migration,invasion and proliferation were investigated by cell scratch test,Transwell test and CCK-8 test.Then the expression of MMP-1,Akt,ERK and TGF-β were detected by ELISA kit to explore the mechanism of PHB-1.Finally,the effect of it on the subcellular structure of cells was observed by transmission electron microscope.Finally,a nude mouse model was constructed to explore the effect of PHB-1 on tumor growth at the animal level.Results:1.SK-MES-1 cells were transfected with plasmid to construct stable cell lines with lower and higher expression of PHB-1,and Real time PCR confirmed that the expression of PHB-1 in SK-MES-1 cells was still stable after cell passage cultivation;2.Higher expression of PHB-1 promoted SK-MES-1 cell proliferation,migration and invasion compared with un-transfected SK-MES-1 cells(Control group);3.Lower expression of PHB-1 inhibited SK-MES-1 cell proliferation,migration and invasion;4.In the supernatant of SK-MES-1 cells overexpressing PHB-1,the expression of AKT and MMP-1 protein was significantly higher than that of the control group,but the expression of TGF-β was down-regulated;The expression of AKT and MMP-1 protein in the cell supernatant of the PHB-1 low expression group was lower than that of the control group,while the expression of TGF-P was significantly up-regulated.There was no significant difference in the expression of ERK in the three groups;5.It was observed by transmission electron microscope that the mitochondria of the cells with lower PHB-1 expression were dissolved and the endoplasmic reticulum expanded,but the subcellular structure of the control and high-expression groups was no significant abnormality;6.The tumor model of nude mice was not constructed successfully.Conclusions:1.PHB-1 overexpression enhanced cell proliferation,and decreased cell proliferation after low expression,which may be related to the activation and inhibition of AKT and TGF-β proteins.2.PHB-1 overexpression enhanced cell migration,and decreased cell migration after low expression,which may be related to the expression of MMP-1 protein.3.The expression of PHB-1 is related to the invasion ability of lung squamous cell carcinoma cells in vitro.4.Inhibition of PHB-1 expression leads to the decrease of SK-MES-1 cell proliferation,migration and invasion ability,which may be related to the changes of mitochondria,endoplasmic reticulum and other subcellular structures.