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Abstract：
We investigated the effects of sevoflurane exposure on the expression of matrix metalloproteinase (MMP), expression and ablation of natural killer group 2, member D (NKG2D) ligands (UL16-binding proteins 1-3 and major histocompatibility complex class I chain-related molecules A/B), and natural killer (NK) cell-mediated cytotoxicity in breast cancer cells.
Three human breast cancer cell lines (MCF-7, MDA-MB-453, and HCC-70) were incubated with 0 (control), 600 (S6), or 1200 μM (S12) sevoflurane for 4 h. The gene expression of NKG2D ligands and their protein expression on cancer cell surfaces were measured using multiplex polymerase chain reaction (PCR) and flow cytometry, respectively. Protein expression of MMP-1 and -2 and the concentration of soluble NKG2D ligands were analyzed using western blotting and enzyme-linked immunosorbent assays, respectively.
Sevoflurane downregulated the mRNA and protein expression of the NKG2D ligand in a dose-dependent manner in MCF-7, MDA-MB-453, and HCC-70 cells but did not affect the expression of MMP-1 or -2 or the concentration of soluble NKG2D ligands in the MCF-7, MDA-MB-453, and HCC-70 cells. Sevoflurane attenuated NK cell-mediated cancer cell lysis in a dose-dependent manner in MCF-7, MDA-MB-453, and HCC-70 cells (P = 0.040, P = 0.040, and P = 0.040, respectively).
Our results demonstrate that sevoflurane exposure attenuates NK cell-mediated cytotoxicity in breast cancer cells in a dose-dependent manner. This could be attributed to a sevoflurane-induced decrease in the transcription of NKG2D ligands rather than sevoflurane-induced changes in MMP expression and their proteolytic activity.
Three human breast cancer cell lines (MCF-7, MDA-MB-453, and HCC-70) were incubated with 0 (control), 600 (S6), or 1200 μM (S12) sevoflurane for 4 h. The gene expression of NKG2D ligands and their protein expression on cancer cell surfaces were measured using multiplex polymerase chain reaction (PCR) and flow cytometry, respectively. Protein expression of MMP-1 and -2 and the concentration of soluble NKG2D ligands were analyzed using western blotting and enzyme-linked immunosorbent assays, respectively.
Sevoflurane downregulated the mRNA and protein expression of the NKG2D ligand in a dose-dependent manner in MCF-7, MDA-MB-453, and HCC-70 cells but did not affect the expression of MMP-1 or -2 or the concentration of soluble NKG2D ligands in the MCF-7, MDA-MB-453, and HCC-70 cells. Sevoflurane attenuated NK cell-mediated cancer cell lysis in a dose-dependent manner in MCF-7, MDA-MB-453, and HCC-70 cells (P = 0.040, P = 0.040, and P = 0.040, respectively).
Our results demonstrate that sevoflurane exposure attenuates NK cell-mediated cytotoxicity in breast cancer cells in a dose-dependent manner. This could be attributed to a sevoflurane-induced decrease in the transcription of NKG2D ligands rather than sevoflurane-induced changes in MMP expression and their proteolytic activity.
摘要：
■我们研究了七氟烷暴露对基质金属蛋白酶（MMP）表达的影响，自然杀伤组2，成员D（NKG2D）配体（UL16结合蛋白[ULBP]1-3和主要组织相容性复合物I类链相关分子[MIC]A/B）的表达和消融，和自然杀伤（NK）细胞介导的乳腺癌细胞的细胞毒性。
■将三种人乳腺癌细胞系（MCF-7，MDA-MB-453和HCC-70）与0（对照）一起孵育，600(S6),或1200μM（S12）七氟醚4h。使用多重聚合酶链反应（PCR）和流式细胞术测量NKG2D配体的基因表达及其在癌细胞表面的蛋白表达，分别。通过蛋白质印迹和酶联免疫吸附试验分析MMP-1和2的蛋白表达和可溶性NKG2D配体的浓度,分别。
■七氟醚在MCF-7、MDA-MB-453和HCC-70细胞中以剂量依赖性方式下调NKG2D配体的mRNA和蛋白表达。然而,它不影响MCF-7，MDA-MB-453和HCC-70细胞中MMP-1和2的表达或可溶性NKG2D配体的浓度。七氟醚在MCF-7，MDA-MB-453和HCC-70细胞中以剂量依赖性方式减弱NK细胞介导的癌细胞溶解（分别为P=0.040，0.040和0.040）。
■我们的结果表明，七氟烷暴露可以以剂量依赖性方式减弱NK细胞介导的乳腺癌细胞的细胞毒性。这可能归因于七氟醚诱导的NKG2D配体转录的减少，而不是七氟醚诱导的MMP表达及其蛋白水解活性的变化。
■将三种人乳腺癌细胞系（MCF-7，MDA-MB-453和HCC-70）与0（对照）一起孵育，600(S6),或1200μM（S12）七氟醚4h。使用多重聚合酶链反应（PCR）和流式细胞术测量NKG2D配体的基因表达及其在癌细胞表面的蛋白表达，分别。通过蛋白质印迹和酶联免疫吸附试验分析MMP-1和2的蛋白表达和可溶性NKG2D配体的浓度,分别。
■七氟醚在MCF-7、MDA-MB-453和HCC-70细胞中以剂量依赖性方式下调NKG2D配体的mRNA和蛋白表达。然而,它不影响MCF-7，MDA-MB-453和HCC-70细胞中MMP-1和2的表达或可溶性NKG2D配体的浓度。七氟醚在MCF-7，MDA-MB-453和HCC-70细胞中以剂量依赖性方式减弱NK细胞介导的癌细胞溶解（分别为P=0.040，0.040和0.040）。
■我们的结果表明，七氟烷暴露可以以剂量依赖性方式减弱NK细胞介导的乳腺癌细胞的细胞毒性。这可能归因于七氟醚诱导的NKG2D配体转录的减少，而不是七氟醚诱导的MMP表达及其蛋白水解活性的变化。
