| Backgrounds:Lung cancer is the first-leading cause of cancer deaths. The effects of traditional chemotherapy have reached a plateau. The molecular targeted therapies have become main methods for advanced non-small cell lung cancer (NSCLC) in recent years. Lung cancer patients Especially with epidermal growth factor receptor (EGFR) mutations, can clearly benefited from EGFR tyrosine kinase inhibitors (TKI). Nevertheless, the majority of patients have experienced clinical drug resistance after treated for some time. Drugs targeting EGFR receptors combined with cytokine-induced killer (CIK) cells in anti-tumor treatment have attracted much attention in recent years. It will help provide new ideas for the treatment of NSCLC to study the effects of EGFR-TKI on CIK cells cytotoxicity against human lung adenocarcinoma cell line HCC827 and its molecular mechanism.Objective:To observe the effects of epidermal growth factor receptor tyrosine kinase inhibitor (EGFR TKI) erlotinib and EGFR downstream regulators on the expression of natural killer group 2, member D receptor (NKG2D) ligands on human lung adenocarcinoma cancer HCC827 cells and detect the cytotoxicity mediated by cytokine-induced killer (CIK) cells against HCC827 cells after treated with erlotinib.Methods:1) After the HCC827 cells are processed by the erlotinib on final concentration of 5, 10μ·L-1, the change of the expression of NKG2D ligands major histocompatibility complex class I chain-related protein (MIC) A, B and UL16 binding protein (ULBP) 1,2,3 on HCC827 cells was detected by flow cytometery.2) After cultured 24h and treated with the inhibitors of phosphatidylinositol 3-kinase LY294002, the inhibitors of mitogen-activated protein kinases SB203580, the inhibitors of signal transducers and activators of transcription 3 STAT21, and the inhibitors of protein kinase C Rottlerin respectively, the expression of NKG2D ligands (MICA, MICB, ULBP1, ULBP2, ULBP3) was assayed by flow cytometery again, then was compared with the blank control group.3) Cytotoxicities of CIK cells against HCC827 cells in the blank control group and erlotinib group, and cytotoxicities of CIK cells blocked with NKG2D monoclonal antibody at effector-to-target cell ratio of 20:1 against HCC827 cells in both groups, were detected by LDH releasing ass_ay.Results:1) Compared with the blank control group, the expression of MICB and ULBP1 were increased notably (P<0.05) and MICA, ULBP2 and ULBP3 were not changed significantly (P>0.05) on HCC827 cells in the erlotinib 5, 10μmol·L-1 group. The expression of NKG2D ligands (MICA, MICB, ULBP1, ULBP2, ULBP3) was no difference in the erlotinib 5μmol·L-1 group and 10μmol·L-1 group (P>0.05). It indicated that erlotinib hydrochloride can upregulate the expression of NKG2D ligands on human lung adenocarcinoma HCC827 cells with EGFR mutations.2) Compared with the blank control group, the expression of NKG2D ligands (MICA, MICB, ULBP1, ULBP2, ULBP3) in the SB203580 group was not significantly different CP>0.05), the expression of MICA and MICB in the STAT21 group and ULBP1 in the Rottlerin group were obviously higher (P<0.05), and the expression of MICA in the LY294002 group was lower (P<0.05). The killing activity of CIK cells to HCC827 cells in the erlotinib group was significantly stronger than the blank control group (P<0.05) at the same effector -to-target cell ratio. It indicated that the downstream regulators of EGFR were involved in regulating the expression of NKG2D ligands on HCC827cells. It can affecte the expression of NKG2D ligands on HCC827 cells to suppress these regulators.3) At the effector -to-target cell ratio of 20:1, the killing activity of CIK cells blocked by NKG2D monoclonal antibody to HCC827 cells between the blank control group and erlotinib 10μmol·L-1 group showed no significance difference. It indicated that erlotinib hydrochloride can enhanced the killing activity of CIK to human lung adenocarcinoma HCC827 cells with EGFR mutations.Conclusion:Erlotinib hydrochloride can enhanced the killing activity of CIK to human lung adenocarcinoma HCC827 cells with EGFR mutations by upregulating the expression of NKG2D ligands. |
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