羟考酮对人结肠癌细胞转移能力的影响：μ、κ受体在其中的作用

Effect of oxycodone on migration of human colon cancer cells： the role of μ and κ receptors

目的评价羟考酮对人结肠癌细胞转移能力的影响及μ、κ受体在其中的作用。方法人结肠癌HCT116细胞，以1×106个/ml的密度接种于24孔板或6孔板，0.5 ml/孔或2 ml/孔，144个培养孔，采用随机数字表法，将其分为6组（n＝24）：对照组（C组）、1、5、10 μmol/L羟考酮组（O1组、O2组和O3组）、羟考酮＋ μ受体拮抗剂CTOP组（O2＋CTOP组）、羟考酮＋κ受体拮抗剂nor-binaltorphimine组（O2＋BNI组）。O1组、O2组和O3组分别采用相应浓度羟考酮孵育，O2＋CTOP组和O2＋BNI组分别采用5 μmol/L羟考酮＋20 μmol/L CTOP或5 μmol/L羟考酮＋nor-binaltorphimin 20 μmol/L孵育24 h。测定侵袭细胞数、迁移细胞数、小G蛋白A（RhoA）、小G蛋白Rho相关激酶-1（ROCK1）、基质金属蛋白酶-2（MMP-2）和MMP-9的水平。结果与C组比较，O1组、O2组和O3组侵袭细胞数和迁移细胞数依次降低，细胞RhoA、ROCK1、MMP-2和MMP-9的水平依次降低（P〈0.05），O2＋BNI组上述指标比较差异无统计学意义（P〉0.05）；与O2组比较，O2＋BNI组侵袭细胞数和迁移细胞数增加，细胞RhoA、ROCK1、MMP-2和MMP-9的水平升高（P〈0.05），O2＋CTOP组细胞上述指标比较差异无统计学意义（P〉0.05）。结论羟考酮可抑制人结肠癌细胞转移能力，其机制全部与激活κ受体后抑制RhoA/ROCK1通路激活有关，而与μ受体无明显关系。

Objective To evaluate the effect of oxycodone on migration of human colon cancer cells and the role of μ and κ receptors.Methods The human colon cancer HCT116 cells at the logarithmic growth phase were seeded in 24-well or in 6-well plates at a density of 1×106 cells/ml（0.5 ml/well or 2 ml/well, 144 wells in total）. The cells were divided into 6 groups（n=24 each）using a random number table： control group（group C）, 1, 5 and 10 μmol/L oxycodone groups（group O1, group O2 and group O3）, oxycodone plus μ receptor antagonist CTOP group（group O2＋ CTOP）and oxycodone plus κ receptor antagonist nor-binaltorphimine group（group O2＋ BNI）. The cells were incubated for 24 h with oxycodone 1, 5 and 10 μmol/L in O1, O2 and O3 groups, respectively.The cells were incubated for 24 h with 5 μmol/L oxycodone plus 20 μmol/L CTOP and 5 μmol/L oxycodone plus nor-binaltorphimin 20 μmol/L in O2＋ CTOP and O2＋ BNI groups, respectively.The invaded and migrated cells were counted, and the levels of Ras homolog gene family member A（RhoA）, Rho-associated protein kinase 1（ROCK1）, matrix metalloproteinase-2（MMP-2）and MMP-9 were detected.Results Compared with group C, the number of invaded and migrated cells was gradually decreased, and the levels of RhoA, ROCK1, MMP-2 and MMP-9 were gradually decreased in O1, O2 and O3 groups（P〈0.05）, and no significant change was found in the parameters mentioned above in group O2＋ BNI（P〉0.05）. Compared with group O2, the number of invaded and migrated cells was significantly increased, and the levels of RhoA, ROCK1, MMP-2 and MMP-9 were increased in group O2＋ BNI（P〈0.05）, and no significant change was found in the parameters mentioned above in group O2＋ CTOP（P〉0.05）.Conclusion Oxycodone can inhibit the migration of human colon cancer cells, and the mechanism is totally related to inhibition of RhoA/ROCK1 signaling pathway activation after activating κ receptors, but not related to μ receptors.