关键词: <, p>, 肝组织, 肝癌组织, 原代组织培养, 成年小鼠<, /p>

Abstract: Abstract: To investigate the influence of primary culture on physiochemical properties of normal hepatic tissues and hepatic tumor tissues of adult mice． The hepatic tumor tissues ( T) and tumor adjacent tissues ( P) of 9 month-aged H-ras12V transgenic mice and normal hepatic tissues ( Wt) of 9 month-aged C57 /BL6J mice were collected to perform the in vitro primary culture． The morphological changes of cultured hepatocytes were detected by hematoxylin-eosin staining during 12 days． The whole-protein variation tendency of cultured hepatic tissues were detected by bradford staining during 48 h． The protein expression levels of p-EＲK，p-AKT，p-mTOＲ，Cyclin D1，and GAPDH of cultured hepatic tissues were detected by western blot during 48 h． The mＲNA expression levels of Cyclin D1，GAPDH，UQCＲB，and 18s ＲNA of cultured hepatic tissues were detected by ＲT-qPCＲ during 48 h．The survival condition of primary cultured hepatocytes of Wt，P，and T were fine during 6 days and the morphological changes were resemble． After 6 days，the cell number of hepatocytes of Wt and P decreased gradually． However，hepatocytes of T dead quickly and appeared a few clone-like cell masses． The whole-protein variation tendencies in Wt，P，and T were resemble． The abundance of protein bands in 130 ～ 170 kμ increased first and then decreased during 48 h． The abundance of protein bands in 40 ～ 130 kμ aincreased during 48 h． The protein expression levels of p-EＲK，p-AKT，p-mTOＲ，Cyclin D1，and GAPDH in primary cultured hepatic tissues decreased gradually during 48 h． The mＲNA expression levels of Cyclin D1 in hepatic tissues of Wt，P，T and GAPDH in Wt，P increased during 48 h． The mＲNA expression levels of GAPDH in hepatic tissues of T and UQCＲB in Wt，P，T decreased first and then increased during 48 h． There is no regularity variation tendency for the expression levels of 18s ＲNA． During the short-term primary culture for hepatic tissues and hepatic tumor tissues of adult mice，although morphology changes of hepatocytes are not obviously different，the mＲNA and protein expression levels of many geneschange significantly． These changes will give great influences on the result of primary culture related experiments．

Key words: <p>hepatic tissue, hepatic tumor tissue, primary tissue culture, adult mice</p>