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目的研究罗勒多糖对肝癌细胞MHCC97H和MHCC97L缺氧模型组蛋白去甲基化酶LSD1、JMJD2B及JARID1B表达的影响,探讨罗勒多糖抗肿瘤侵袭转移作用与表观遗传修饰的关系。方法采用二氯化钴诱导肝癌细胞缺氧,建立肝癌细胞MHCC97H和MHCC97L体外缺氧模型,通过不同剂量罗勒多糖干预24h,细胞划痕实验检测缺氧条件下肝癌细胞的迁移能力,实时荧光定量PCR法检测各组肝癌细胞中HIF-1α、LSD1、JMJD2B及JARID1B mRNA表达水平,Westernblot法检测各组肝癌细胞中HIF-1α、LSD1、JMJD2B及JARID1B蛋白表达情况。结果罗勒多糖能抑制缺氧条件下MHCC97H和MHCC97L细胞的迁移能力,下调细胞中HIF-1αmRNA及蛋白的表达水平,与缺氧模型组相比有显著性差异(P<0.05);罗勒多糖能够下调MHCC97H细胞缺氧条件下LSD1、JMJD2B、JARID1B mRNA和蛋白的表达,与缺氧模型组相比有显著性差异(P<0.05);缺氧条件下MHCC97L细胞中LSD1、JMJD2B、JARID1B mRNA和蛋白的表达与正常对照组无显著性差别(P>0.05),罗勒多糖对这三种酶的表达亦无显著作用。结论罗勒多糖能改善不同转移潜能肝癌细胞MHCC97H和MHCC97L的缺氧微环境,抑制肿瘤细胞的侵袭转移。其中罗勒多糖对高转移潜能肝癌细胞MHCC97H的抗侵袭转移作用与其对细胞中组蛋白去甲基化酶LSD1、JMJD2B、JARID1B的调节有关,而对低转移潜能肝癌细胞MHCC97L侵袭转移能力的调节未发现与LSD1、JMJD2B、JARID1B有明显关系。
Objective To study the effects of basilain on the expressions of histone demethylase LSD1, JMJD2B and JARID1B in hepatocarcinoma cells MHCC97H and MHCC97L hypoxia model and to explore the relationship between the anti-tumor invasion and metastasis of basil polysaccharide and epigenetic modification. Methods HCC cells were induced to hypoxia by using cobalt chloride chloride. Hypoxic models of MHCC97H and MHCC97L cells were established in vitro. Interfering with different doses of basil, the migration of HCC cells under hypoxia was detected by cell scratch assay. Real-time fluorescence quantitative PCR The expressions of HIF-1α, LSD1, JMJD2B and JARID1B mRNA in hepatocellular carcinoma cells were detected by Western blot. The protein expressions of HIF-1α, LSD1, JMJD2B and JARID1B were detected by Western blot. Results Basil polysaccharide could inhibit the migration of MHCC97H and MHCC97L cells under hypoxia, down-regulate the expression of HIF-1αmRNA and protein in Hypoxia model group (P <0.05), and basil polysaccharide can down-regulate The expressions of LSD1, JMJD2B and JARID1B mRNA and protein in MHCC97H cells under hypoxia were significantly different from those in hypoxia model group (P <0.05). The mRNA and protein expression of LSD1, JMJD2B and JARID1B in MHCC97L cells under hypoxia There was no significant difference between the two groups (P> 0.05). The basal polysaccharide had no significant effect on the expression of these three enzymes. Conclusion basil polysaccharide can improve the hypoxia microenvironment of MHCC97H and MHCC97L cells with different metastatic potentials and inhibit the invasion and metastasis of tumor cells. The anti-invasion and metastasis of basilar polysaccharides on highly metastatic potential hepatocarcinoma cell line MHCC97H was related to the regulation of histone demethylase (LSD1), JMJD2B and JARID1B in cell lines, but no effect on the invasion and metastasis of low metastatic potential hepatocarcinoma cell line MHCC97L Significantly related to LSD1, JMJD2B, JARID1B.