The Effect Of Conditional Knockout LKB1/PTEN Combination On The Occurrence Of Liver Tumors In Mice

Cells accumulate the genetic and epigenetic changes in the physiological activity, leadingthe cells to grow into a non-controlled state by changing the proteome of the cells, which isCalled cancer. In recent years, The incidence of cancer has been on the rise, and has become aserious threat to the human life and health. In China, hepatocellular liver cancer (hepatocellularcarcinoma, HCC) is the second leading cause of cancer death, and also the fifth most commoncancer worldwide.There is a variety of factors related to HCC pathogenesis. including viral infections,hepatitis, aflatoxin, alcohol, obesity, diabetes, and primarily regulated by oncogenes and tumorsuppressor genes. myc and ras are the oncogenes associated with the incidence of HCC,andtumor suppressor genes include p53, rb, runx3, pten and so on. HCC incidence is the result of thecombined effects of these factors.The mutation rate of pten is high in HCC, approximately40%.pten gene was discovered in1997, considered to be an important tumor suppressor gene following rb and p53. pten candephosphorylate PIP3to generate PIP2, negativly regulating the PI3K/AKT signalingpathways,and inhibit tumor growth at last.In some cancers, we can’t detect the oncogene mutation.such as retinoblastoma,only rbgene mutation is detected; In mice, the inactivation of p53and rb gene can cause small cell lungcancer. and this prompted us, in some cases, the tumor suppressor gene mutation can lead totumor alonely.Given the high mutation rate of pten in HCC and the key role of lkb1/pten inPI3K/AKT/mTOR signaling pathway,we propose to esplore the effect of lkb1/pten inactivationto liver.In this paper, we use the Cre-loxP system.we knockout lkb1/pten Specifically in the liverby intravenous injection, and then study the effect of lkb1/pten combination inactivation to theliver. The mice will be a month old when lkb1/pten were knockouted.when six months after theexperiment, the liver tumor nodes of the mouse can be found and further to be verified by HEstaining and the marker for HCC. We study the expression of protein associated with livertumors by western and immunohistochemistry.With lkb1/pten inactivation leading to HCC in mice,we then want to look at the effect oncells,and study the relevant mechanisms. This paper selected3T3(mouse fibroblasts) andAML12(normal mouse liver cells) two cell lines. We obtain the desired cell lines by buildingshlkb1shPTEN lentiviral plasmids, viral packaging, lentivirus transfecting and puromycin blasticidin antibiotic selection. We study the expression of protein associated with liver tumorsby western.In this article, western detection results are not satisfactory, and the change of protein isdifferent in mouse liver tumor tissue and cells.it’s may be due to the low efficiency ofknockdown in cells, and also may be due to the presence of different mechanisms. Therefore, themechanism of lkb1/pten inactivation leading to HCC in mice is required to be further study...