Department of Pediatric Surgery, Zhujiang Hospital, Southern Medical University, Guangzhou, Guangdong, China.
Department of Thyroid Hernia Vascular Pediatric Surgery, The Second People's Hospital of Foshan, Foshan, Guangdong, China.
J Cell Physiol. 2024 Sep;239(9):e31402. doi: 10.1002/jcp.31402. Epub 2024 Aug 7.
The pathogenesis of Hirschsprung's disease (HSCR) is complex. Recently, it has been found that histone modifications can alter genetic susceptibility and play important roles in the proliferation, differentiation and migration of neural crest cells. H3K36 methylation plays a significant role in gene transcriptional activation and expression, but its pathogenic mechanism in HSCR has not yet been studied. This study aimed to elucidate its role and molecular mechanism in HSCR. Western blot analysis, immunohistochemistry (IHC) and reverse transcription-quantitative PCR (RT‒qPCR) were used to investigate H3K36 methylation and methyltransferase levels in dilated and stenotic colon tissue sections from children with. We confirm that SMYD2 is the primary cause of differential H3K36 methylation and influences cell proliferation and migration in HSCR. Subsequently, quantitative detection of m6A RNA methylation revealed that SMYD2 can alter m6A methylation levels. Western blot analysis, RT-qPCR, co-immunoprecipitation (co-IP), and immunofluorescence colocalization were utilized to confirm that SMYD2 can regulate METTL3 expression and affect m6A methylation, affecting cell proliferation and migration. These results confirm that the H3K36 methyltransferase SMYD2 can affect cell proliferation and migration in Hirschsprung's disease by regulating METTL3. Our study suggested that H3K36 methylation plays an important role in HSCR, confirming that the methyltransferase SMYD2 can affect m6A methylation levels and intestinal nervous system development by regulating METTL3 expression.
先天性巨结肠症（HSCR）的发病机制较为复杂。最近发现，组蛋白修饰可以改变遗传易感性，并在神经嵴细胞的增殖、分化和迁移中发挥重要作用。H3K36 甲基化在基因转录激活和表达中起重要作用，但在 HSCR 中的致病机制尚未得到研究。本研究旨在阐明其在 HSCR 中的作用和分子机制。通过 Western blot 分析、免疫组织化学（IHC）和逆转录-定量 PCR（RT-qPCR），我们研究了儿童扩张段和狭窄段结肠组织切片中的 H3K36 甲基化和甲基转移酶水平。我们证实 SMYD2 是导致 H3K36 甲基化差异的主要原因，并影响 HSCR 中的细胞增殖和迁移。随后，对 m6A RNA 甲基化的定量检测表明 SMYD2 可以改变 m6A 甲基化水平。Western blot 分析、RT-qPCR、共免疫沉淀（co-IP）和免疫荧光共定位实验证实 SMYD2 可以调节 METTL3 的表达并影响 m6A 甲基化，从而影响细胞增殖和迁移。这些结果证实 H3K36 甲基转移酶 SMYD2 可以通过调节 METTL3 影响 HSCR 中的细胞增殖和迁移。我们的研究表明 H3K36 甲基化在 HSCR 中起重要作用，证实甲基转移酶 SMYD2 可以通过调节 METTL3 表达来影响 m6A 甲基化水平和肠道神经系统发育。
