中文摘要：

目的：采用重组慢病毒系统，通过过表达和 RNA 干涉的方法分别增加和降低 NDRG2（N - Myc down-stream regulated gene 2）基因后，检测人结肠癌细胞系 DLD -1中丙酮酸脱氢酶（PDH）的表达水平及氧消耗率（OCR）。方法：分别采用表达 NDRG2慢病毒载体 pLenti6- NDRG2和 NDRG2 shRNA 慢病毒载体 pLKO.1-shNDRG2制备病毒载体并感染 DLD -1细胞，经两周耐药筛选，获得稳定感染的细胞株。采用 Real - time PCR 和 Western blot 的方法明确 NDRG2在稳定感染细胞株中的表达情况，再使用上述方法检测稳定感染细胞株中 PDH 的表达水平，最后，通过 Seahorse 能量呼吸代谢仪，检测稳定感染细胞株中的 OCR。结果：Real -time PCR 和 Western blot 结果表明，在慢病毒 pLenti6- NDRG2稳定感染的 DLD -1细胞中，NDRG2表达上调，PDH 表达上调；pLKO.1- shNDRG2稳定感染的 DLD -1细胞中，NDRG2的表达下调，PDH 表达下调。Seahorse 结果表明，NDRG2过表达细胞株中细胞的 OCR 上升，而 NDRG2 RNA 干涉细胞株中细胞的 OCR 明显下降。结论：通过过表达和抑制 NDRG2基因，证实了 NDRG2作为抑癌候选基因能够有效促进结肠癌细胞系 DLD -1中丙酮酸脱氢酶 PDH 的表达，并促进有氧氧化。

英文摘要：

Objective：To investigate the effects of over - expression and knockdown of NDRG2（N - Myc down-stream regulated gene 2）gene on the expression of pyruvate dehydrogenase（ PDH）and oxygen consumption rate （OCR）of human colorectal carcinoma DLD - 1 cells by using recombinant lentivirus vectors containing NDRG2 or NDRG2 shRNA. Methods：The lentiviruses containing NDRG2 or NDRG2 shRNA,prepared separately from NDRG2 lentiviral expression vector pLenti6 - NDRG2 or NDRG2 shRNA lentiviral expression vector pLKO. 1 - shNDRG2, were used to infect human colorectal carcinoma cell line DLD - 1. The stable cell lines were obtained by selection with blasticidin or puromycin for 2 weeks. Real - time PCR and Western blot were employed to examine the expression of NDRG2 and PDH in these stable cell lines. Seahorse Extracellular Flux（XF24e ）analyzer was used to determine basal oxygen consumption rate（OCR）of these stable cell lines. Results：The mRNA and protein levels of NDRG2 were up -regulated in DLD - 1 cell lines infected with pLenti6 - NDRG2,and down - regulated in those with pLKO. 1 -shNDRG2. The PDH and OCR were up - regulated in DLD - 1 cell lines stably infected with pLenti6 - NDRG2,and down - regulated in those with pLKO. 1 - shNDRG2. Conclusion：NDRG2,as a candidate tumor suppressor gene, could promote the expression of PDH and boost oxidative phosphorylation.