LYRM2表达通过调控糖酵解代谢影响结肠癌RKO细胞增殖、侵袭

LYRM2 regulates the proliferation and invasion of colon cancer RKO cells by regulating glycolytic metabolism

目的探讨结肠癌中LYRM2表达对结肠癌RKO细胞增殖、迁移、侵袭的影响及潜在机制。方法利用LYRM2过表达载体L2HA和干扰载体SH1KD构建稳转结肠癌细胞系,CCK-8检测细胞增殖活力,划痕实验、Transwell实验检测细胞迁移、侵袭能力;Seahorse法检测细胞糖酵解代谢及乳酸生成量,试剂盒法检测细胞糖酵解代谢限速酶活性。结果与对照组细胞相比,LYRM2过表达组RKO细胞的LYRM2表达水平[(167.10±3.73)vs(8.79±0.17),P<0.001]、增殖第6天细胞吸光度[OD450 nm:(1.83±0.08)vs(1.20±0.09),P<0.001]、48 h后迁移距离[(46.67±0.33)μm vs(28.33±0.33)μm,P<0.001]、侵袭细胞数目[(941.70±39.55)vs(469.30±21.53),P<0.001]、糖酵解代谢(包括细胞内ATP含量、乳酸生成)及细胞内HK2活性[(0.126±0.002)U vs(0.098±0.002)U,P<0.001]均显著升高;LYRM2敲低组RKO细胞的LYRM2表达水平[(1.13±0.11)vs(11.51±0.82),P<0.01]、第6天细胞吸光度[OD450 nm:(0.82±0.07)vs(1.35±0.09),P<0.001]、48 h后迁移距离[(3.67±0.33)μm vs(27.67±0.33)μm,P<0.001]、侵袭细胞数目[(87.00±9.29)vs(504.30±3.48),P<0.001]、糖酵解代谢及细胞内HK2活性[(0.076±0.002)U vs(0.099±0.002)U,P<0.001]均明显抑制。结论LYRM2过表达促进结肠癌RKO细胞的增殖、迁移及侵袭能力,其通过增强HK2活性促进RKO细胞糖酵解代谢,有望成为结肠癌治疗的新靶点。

Purpose To investigate the effect of LYRM2 on the proliferation and migration of colon cancer RKO cells and its underlying mechanism.Methods The stable colon cancer cell lines were established by LYRM2 overexpression vector L2HA and interference vector SH1KD.CCK-8,scratch test and transwell chamber test were used to detect cell proliferation,migration and invasion.Seahorse method was used to detect the glycolytic metabolism and lactic acid production of cells,and kit method was used to detect the activity of rate limiting enzyme of glycolytic metabolism.Results Compared with control cells,the LYRM2 expression level[(167.10±3.73)vs(8.79±0.17),P<0.001],cell absorbance on the 6th day[OD450 nm:(1.83±0.08)vs(1.20±0.09),P<0.001],migration distance after 48 h[(46.67±0.33)μm vs(28.33±0.33)μm,P<0.001],number of invasive cells(941.70±39.55)vs(469.30±21.53),P<0.001],glycolytic metabolism(including intracellular ATP and lactate production)and intracellular HK2 activity[(0.126±0.002)U vs(0.098±0.002)U,P<0.001]of LYRM2 overexpressing RKO cells were significantly increased.The LYRM2 expression level[(1.13±0.11)vs(11.51±0.82),P<0.01],cell absorbance on day 6[OD450 nm:(0.82±0.07)vs(1.35±0.09),P<0.001],migration distance after 48 h[(3.67±0.33)μm vs(27.67±0.33)μm,P<0.001],number of invasive cells[(87.00±9.29)vs(504.30±3.48),P<0.001],glycolytic metabolism and intracellular HK2 activity[(0.076±0.002)U vs(0.099±0.002)U,P<0.001]of LYRM2 knockdown RKO cells were significantly inhibited.Conclusion LYRM2 overexpression can promote the proliferation,migration and invasion of RKO cells.It promotes the glycolysis metabolism of RKO cells by enhancing the activity of HK2,which is expected to become a potential new target for the treatment of colon cancer.