乳酸脱氢酶C_4在早期邪毒壅盛证荷瘤小鼠肿瘤组织中的表达及意义

Expression and Significance of Lactate Dehydrogenase C_4 in Tumor Tissues of H_(22) Mice with Obstruction of Evil Syndromes

目的探讨乳酸脱氢酶C4(LDH-C4)在邪毒壅盛证荷瘤小鼠肿瘤组织的表达水平及其意义。方法采用Affy-metrix Gene Chip Mouse Exon 1.0 ST Array及其方法检测乳酸转运和氧化相关基因在H22荷瘤小鼠早期邪毒壅盛、早期气虚、中期阳气虚及中晚期气阴阳虚证肿瘤组织基因表达的差异,芯片重复两次;并采用实时荧光定量PCR对两种高乳酸亲和力的脱氢酶基因Ldhb和Ldhc在各种证候中的表达量进行检测。结果乳酸转运相关基因的表达要高于氧化相关基因,尤其以H22接种后早期最明显;Ldhb基因的表达在H22接种后早期、中期和晚期变化不明显,而Ldhc基因在H22接种后早期气虚、中期和晚期的表达也不明显,且与Ldhb表达量相近,但它在早期邪毒壅盛证荷瘤小鼠肿瘤组织中表达量显著升高。结论在荷瘤小鼠肿瘤组织中,尤其是H22接种后早期,肿瘤细胞产生的乳酸作用不仅在于氧化产能;LDH-C4可能代替LDH-B4参与了早期邪毒壅盛证荷瘤小鼠肿瘤组织缺氧肿瘤细胞的代谢拯救途径。

Objective To investigate the expression and significance of lactate dehydrogenase C4 in tumor tissues of H22 mice with obstruction of evil syndromes in early stage. Methods Expression of genes related to lactate transport and oxidation in the tumor tissues of H22 mice with syndromes of obstruction of evil syndromes,＂qi＂ deficiency,deficiency of ＂qi＂ and yang,or deficiency of ＂qi＂,＂yin＂ and ＂yang＂,were analyzed by Affymetrix GeneChip Mouse Exon 1.0 ST Array with two repeats,and those of Ldhb and Ldhc were examined by real-time quantitative Polymerase chain reaction（PCR）. Results Higher expression of genes related to lactate transport than those related to lactate oxidation in the tumor tissues of H22 mice was observed,especially with syndromes of obstruction of evil syndromes and ＂qi＂ deficiency.Expression of Ldhb had no significant change after H22 tumor cells were vaccinated,but Ldhc had higher expression in tumor tissues of H22 mice with obstruction of evil syndromes in early stage,compared with expression of Ldhb,and those of Ldhc in tumor tissues of H22 mice with ＂qi＂ deficiency,deficiency of ＂qi＂ and ＂yang＂,or deficiency of ＂qi＂,＂yin＂ and ＂yang＂. Conclusion Lactate released by tumor cells can be oxidated to produce energy in tumor tissues of H22 mice,especially in early stage.LDH-C4 may be involved in a metabolic rescue pathway instead of LDH-B4 in hypoxic tumor cells of H22 mice with obstruction of evil syndromes in early stage.