抑制 KLF8 基因对 CD133+肝癌干细胞亚群比例、裸鼠体内成瘤和肺转移能力的影响

Objective To investigate the effects of inhibition of K.LF8 expression on the proportion of CD133+ phenotype cell subsets in the hepatocellular carcinoma stern cells, tumor formation in vivo and lung metastasis in the nude mice. Methods Liver cancer stem cell subsets (LCSCs) with CD133+ phenotype in the lung metastases of liver cancer patients were isolated and cultured. LCSCs were infected with the recombinant lentiviral plasmid (sh-KLF8) interfered by RNA and its negative control empty vector plasmid (sh-control) , respectively. CD133+ phenotype liver cancer stem cell line (LCSC-ssh-KLF8) and control cell line (LCSCssh-eontrol) with stably knocking out K.LF8 gene were constructed, then we constructed the liver cancer stem cell xenograft model and lung metastasis model. The qRT-PCR was used to detect the expression of K.LF8 in the two cell lines, and the ratio of CD133+ phenotypic cells in the two cell lines was detected by flow cytometry. We injected LCSCssh-KLF8 and LCSCssh-eontrol to construct the nude mouse xenograft model of liver cancer stem cells, and observed the effect of knocking out of K.LF8 gene on the tumorigenic ability of liver cancer stem cells in nude mice. We injected LCSCssh-KLF8 and LCSCssh-eontrol into the nude mice through the tail vein to construct the nude mouse lung metastasis model, and observed the effect of knocking out of K.LF8 gene on the lung metastasis ability of liver cancer stem cells in nude mice. K.LF8 protein was detected by immunohistoehemistry in the lung metastases formed by LCSCssh-KLF8 and LCSCssh-eontrol. Results Compared with LCSCssh-eontrol, the relative expression level of K.LF8 mR-JNA in LCSCssh-KLF8 significantly decreased (P<0.05) , and the proportion of CD133+ phenotype stem cell subsets significantly decreased, and the tumor weight and number of lung metastases in vivo significantly decreased in the nude mice, and the positive staining of K.LF8 protein in the lung metastases was much less (all P < 0. 05). Conclusion Inhibiting JVLF8 gene can reduce the proportion of CD133+ phenotype stem cell subsets, tumor formation in vivo, and lung metastasis ability, and therefore KCFS may be the key molecule that inhibits the development and progression of hepatocellular carcinoma. [ABSTRACT FROM AUTHOR]