Shao Meng, Zhang Jiayin, Zhang Jiahui, Shi Hui, Zhang Yu, Ji Runbi, Mao Fei, Qian Hui, Xu Wenrong, Zhang Xu
Jiangsu Key Laboratory of Medical Science and Laboratory Medicine, School of Medicine, Jiangsu University, 301 Xuefu Road, Zhenjiang, Jiangsu 212013 China.
Department of Clinical Laboratory Medicine, The Affiliated People's Hospital of Jiangsu University, Zhenjiang, Jiangsu 212002 China.
Cancer Cell Int. 2020 May 24;20:188. doi: 10.1186/s12935-020-01275-y. eCollection 2020.
The stem cell factor SALL4 is reactivated in human cancers. SALL4 plays diverse roles in tumor growth, metastasis, and drug resistance, but its role in tumor metabolism has not been well characterized.
The glycolytic levels of gastric cancer cells were detected by glucose uptake, lactate production, lactate dehydrogenase activity, ATP level, and hexokinase activity. QRT-PCR and western blot were used to detect the changes in the expression of glycolytic genes and proteins. The downstream target genes of SALL4 were identified by microarray. The regulation of hexokinase II (HK-2) by SALL4 was analyzed by luciferase reporter assay and chromatin immunoprecipitation assay. Transwell migration assay, matrigel invasion assay, cell counting assay and colony formation assay were used to study the roles of HK-2 regulation by SALL4 in gastric cancer cells in vitro. The effects of SALL4 on glycolysis and gastric cancer progression in vivo were determined by subcutaneous xenograft and peritoneal metastasis tumor models in nude mice.
SALL4 knockdown inhibited glucose uptake, lactate production, lactate dehydrogenase activity, ATP level and hexokinase activity in gastric cancer cells, and decreased the expression of glycolytic genes and proteins. Microarray analysis showed that SALL4 knockdown affected glycolysis-related pathway. The regulation of HK-2 gene expression by SALL4 was confirmed by luciferase reporter assay and chromatin immunoprecipitation assay. HK-2 knockdown abrogated the promotion of glycolysis by SALL4 in gastric cancer cells, indicating that HK-2 acts as a downstream effector of SALL4. Moreover, HK-2 knockdown reversed the promoting role of SALL4 in gastric cancer cell proliferation, migration and invasion, suggesting that SALL4 drives gastric cancer progression by upregulating HK-2.
SALL4 promotes gastric cancer progression through HK-2-mediated glycolysis, which reveals a new mechanism for the oncogenic roles of SALL4 in cancer.
干细胞因子SALL4在人类癌症中被重新激活。SALL4在肿瘤生长、转移和耐药性中发挥多种作用，但其在肿瘤代谢中的作用尚未得到充分表征。
通过葡萄糖摄取、乳酸产生、乳酸脱氢酶活性、ATP水平和己糖激酶活性检测胃癌细胞的糖酵解水平。采用QRT-PCR和蛋白质印迹法检测糖酵解基因和蛋白质表达的变化。通过微阵列鉴定SALL4的下游靶基因。通过荧光素酶报告基因检测和染色质免疫沉淀检测分析SALL4对己糖激酶II（HK-2）的调控。采用Transwell迁移检测、基质胶侵袭检测、细胞计数检测和集落形成检测研究SALL4对HK-2的调控在体外胃癌细胞中的作用。通过裸鼠皮下异种移植和腹膜转移瘤模型确定SALL4对体内糖酵解和胃癌进展的影响。
SALL4敲低抑制了胃癌细胞的葡萄糖摄取、乳酸产生、乳酸脱氢酶活性、ATP水平和己糖激酶活性，并降低了糖酵解基因和蛋白质的表达。微阵列分析表明，SALL4敲低影响了糖酵解相关途径。荧光素酶报告基因检测和染色质免疫沉淀检测证实了SALL4对HK-2基因表达的调控。HK-2敲低消除了SALL4对胃癌细胞糖酵解的促进作用，表明HK-2是SALL4的下游效应分子。此外，HK-2敲低逆转了SALL4对胃癌细胞增殖、迁移和侵袭的促进作用，提示SALL4通过上调HK-2促进胃癌进展。
SALL4通过HK-2介导的糖酵解促进胃癌进展，这揭示了SALL4在癌症中致癌作用的新机制。
