Histone Methyltransferase G9a Promotes the Development of Renal Cancer through Epigenetic Silencing of Tumor Suppressor Gene SPINK5

Figure 5

Suppression of SPINK5 contributes to the tumor-promoting function of G9a. (a) The expression of proteins of SPINK5, ANKRD1, PTGS1, P53, and RARRES3 was measured. (b) The expression of G9a was inversely correlated with the expression of SPINK5 in human RCC cell lines though data mining. (c) CCK-8 assay was performed to show the proliferation ability of 786-O after knocking down both G9a and SPINK5 (treated for 12, 24, 36, 48, and 72 h). (d, e) Invasive and migratory abilities of shG9a-shSPINK5-786-O cells were measured through transwell assay and wound healing assay. (f) The expression of protein-related EMT and p53 was measured. (g) ChIP was applied to analyze the interaction of G9a protein (H3K9me2 methyl transferase) and promoter sequence of the downstream target SPINK5. Knockdown of G9a significantly diminished the G9a binding and H3K9me2 level at the promoter region of SPINK5 as determined by ChIP assay.

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