Differential gene expression in Hep3B and Hep3B SOX4-/- cells

Overexpression of SOX4 in various kinds of cancers specimen was associated with poor prognosis of patients; however, the role of SOX4 in angiogenesis or tumor microenvironment modulation remains unclear. Therefore the endogenous SOX4 was knockout and the differential gene expression between Hep3B and Hep3B SOX4-/- cells were examined via genechip. We found that the differentially expressed genes, EzH2, a SOX4-associated partner, and CXCL12, were repressed in Hep3B SOX4-/- cells compared with parental Hep3B; these results were further assessed via qRT-PCR in Hep3B SOX4-/- versus Hep3B cells. Endogenous SOX4 knockout in Hep3B cells via the CRISPR/cas9 system that was designed as Hep3B SOX4-/-, the transcriptome of Hep3B and Hep3B SOX4-/- cells were analyzed via an Affymetrix GeneChip™ Human Transcriptome Array 2.0.

SOX4在多种癌症标本中的过表达与患者不良预后密切相关，但目前SOX4在血管生成或肿瘤微环境调控中的作用仍不明确。为此，本研究对内源性SOX4进行敲除，并通过基因芯片（genechip）检测Hep3B细胞与Hep3B SOX4敲除（SOX4-/-）细胞之间的差异基因表达情况。研究发现，与亲本Hep3B细胞相比，Hep3B SOX4-/-细胞中差异表达基因、SOX4相关结合蛋白EzH2以及CXCL12均呈现表达抑制；上述结果进一步通过实时定量逆转录PCR（qRT-PCR）在Hep3B SOX4-/-细胞与亲本Hep3B细胞中得到验证。本研究通过CRISPR/Cas9系统对Hep3B细胞的内源性SOX4进行敲除，构建得到Hep3B SOX4-/-细胞，并采用Affymetrix GeneChip™人类转录组芯片2.0（Affymetrix GeneChip™ Human Transcriptome Array 2.0）对Hep3B细胞与Hep3B SOX4-/-细胞的转录组进行分析。