Opioids Affect The Immunity Of Mice And Malignant Biological Behavior Of The Colon Cancer Cell

ObjectiveOpioids have become the most important analgesic in cancer patient and perioperative period.However there is no consensus about the impact of opioids on the immunity and cancer.Considering the opioids consumption of cancer patient and the chaos in this field,We used the mouse model suffering from colon cancer by subcutaneous injection to research the effects of opioids on the peripheral immune system including the composition and function of lymphocyte,in the meaning while,we used the mouse colon cancer cell MC38 to research the impact of opioids on the malignant biological behavior including proliferation,migration,invasion and chemotaxis,in which further study was made to unveil the position of mu opioid receptor.The purpose of the research is to provide a theoretical basis for the rational use of opioids in the clinical colon cancer patients and pave the way to the more mechanical research about opioids and cancer.Methods Part 1There were 39 mice received subcutaneous injection with colon cancer cell line CT26,which were divided into 4 groups randomly after successful modeling and administrated the equivalent dose of drugs by vein injection in 0.2ml.The oxycodone group was given 5 mg/kg;the sufentanil group was given 5μg/kg;the fentanyl group was given 50μg/kg every 12 hour for continueous 3 days.12 hours after the last administration,blood and the spleen were collected.The proportion of CD3~+T,CD3~+CD4~+T,CD3~+CD8~+T and NK cells in peripheral blood and spleen were analyzed by flow cytometry,as well as the proportion of MDSC and granular MDSC in spleen.Another 43 mice were handled with the same way and the content of TH1/TH2/TH17cytokines(IL-2,IL-4,IL-6,IL-10,IFN-?,TNF-α,IL-17A)in plasma was detected by CBA assay.Part 2Mouse colon cancer cell line MC38,administrated with PBS,fentanyl,sufentanyl and dezocine respectively in three kinds of levels,was tested for characteristic of proliferation by CCK8 assay and migration,invasion and chemotaxis by using Transwell assay.The opioid receptor antagonist naloxone was applied to further study the change in chemotaxis and invasion that were enhanced originally by opioids.Results Part 11.In the peripheral blood,contrast to the saline group,the percentage of CD3~+T cell,CD3~+CD4~+T cell,CD3~+CD8~+T cell and CD49b~+T cell after sufentanyl and fentanyl administration were decreased,but there was not significant difference in oxycodone group;contrast to the oxycodone group,the percentage of CD3~+T cell,CD3~+CD4~+T cell,and CD3~+CD8~+T cell in the sufentanyl group and the fentanyl group were decreased and the percentage of CD49b~+T cell in the fentenyl group was decreased;the latter two groups had no statistical significance;The ratio of CD4/CD8 among groups had no statistical significance,too.In the spleen,contrast to the saline group and the fentenyl group,the proportion of CD3~+T,CD3~+CD4~+T and CD3~+CD8~+T cells in oxycodone group and sufentanil group was increased.The proportion of CD3~+T lymphocyte and CD3~+CD8~+T lymphocyte in oxycodone group was higher than those in fentanyl group;The ratio of CD4/CD8 among groups had no statistical significance;compared to the saline group,the proportion of NK cell in opioid groups was lower.2.In peripheral blood,compared to the saline group,the concentration of TNF-αin opioid groups was lower and the concentration of TNF-αin oxycodone group and sufentanyl group was higher than that in fentanyl group;compared to the saline group,the concentration of IL-10 in oxycodone group and sufentanyl group was higher;compared to other groups,the ratio of Th1/Th2 in fentanyl group was lower significantly and presented an imbalance of TH1/TH2 to TH2 type immunity.The concentration of IFN-α、IL-6、IL-17A among groups had no statistical significance.The concentration of IL-2 and IL-4 was so little that it could’t be detected.Part 21.In the CCK8 proliferation experiment,the proliferation in opioid groups didn’t have obvious discrepancy with that in the saline group,but there was a significant difference between the 1ng/ml sufentanyl group and the 10ng/ml fentanyl group.2.MC38 treated with the fentanyl,sufentanil and dezocine all had a concentration-dependent up-regulation in migration and invasion in a certain concentration range,except the migration in dezocine group.In low concentrations,MC38 invasion in sufentanyl group presented the strongest among the three opioid groups,followed by the dezoxine group and the migration assay presented no difference;in moderate concentrations,MC38 migration and invasion in sufentanyl group both presented the strongest,followed by the dezocine group;in high concentrations,MC38 invasion in sufentanyl group and dezocine group showed stronger than that in the fentanyl group and the migration assay showed no difference.The invasion increased by opioids in the high concentrations could not be neutralized by the preculture of naloxone.3.The opioids all had an evident effect on the chemotaxis of MC38,which could be partially neutralized by the naloxone in the dezocine group.In low concentrations,MC38 chemotaxis in the sufentanyl group showed the strongest among the three opioid groups,followed by the dezocine group;in moderate concentrations,MC38 chemotaxis in dezocine group showed the strongest,followed by the sufentanyl group and fentanyl group.Conclusions1.In mice inoculated with colon cancer cells,opioids can weaken the immunity in the peripheral blood.Among them,the oxycodone group has little effect on the immunity in compared with the sufentanyl group and the fentanyl group.2.Cell experiments showed that opioids can intensify the migration,invasion and chemotaxis of MC38,but not proliferation.In contrast to the equivalent dose of fentanyl and dezocine,Sufentanyl showed the strongest in promote the malignant biological behavior.This reinforce on invasion can’t be conducted by opioid receptor,but the increase in chemotaxis can be conducted by the opioid receptor partially.