高压纳秒脉冲电场杀伤化疗抗性肿瘤细胞的实验机理研究
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摘要
肿瘤严重危害人类生命和健康,化疗抵抗是当前临床肿瘤治疗所面临的主要屏障之一。为探究高压纳秒脉冲电场是否能够对化疗抵抗的肿瘤细胞具有同等敏感性,以肺癌细胞系A549及顺铂化疗抵抗的A549/R为研究对象,用80个脉冲宽度为200 ns,场强为5～15 kV/cm的纳秒脉冲作用后分析对比其活性情况、致死效应及消融阈值差异。结果表明:当场强为8、10、15 kV/cm纳秒脉冲电场作用下A549/R细胞的存活率分别为5.65%、7.78%、2.80%,显著低于A549(分别为48.31%、26.8%、5.96%);诱导的凋亡和坏死率也显著高于A549(P<0.05);在80个宽度为200 ns,场强为15 kV/cm的脉冲作用下,A549/R单层细胞消融面积是A549的1.59倍;A549/R的消融阈值为～8 kV/cm,显著低于A549(～13 kV/cm)。因此,纳秒脉冲电场为可优先选择杀伤化疗抵抗的A549/R细胞,诱导更高的凋亡率、消融面积,并且有更低的消融阈值。高压纳秒脉冲对化疗抵抗的肿瘤细胞的选择性作用,可望作为一种新型的物理选择方式联合其他肿瘤治疗方法,克服肿瘤治疗临床难点,为化疗抵抗或高恶性度的肿瘤消融提供新策略。
Tumor seriously endanger the lives and health of human people. Chemoresistance is one of the main barriers to successful tumor treatment. In order to explore whether high voltage nanosecond pulsed electric fields can be equally sensitive to chemotherapy-resistant tumor cells compared with its homology tumor cells, lung cancer cell line A549 and cisplatin chemotherapy-resistant A549/R were used as the research object in this paper. After exposing to the 80 nanosecond pulses with a pulse width of 200 ns and a field strength of 5-15 kV/cm, the activity of cell, apoptosis and necrosis and ablation effect were analyzed and compared in groups. The results showed that the survival rate of A549/R was 5.65%, 7.78% and 2.80% respectively when exposed to pulses electric field with strength of 8 kV/cm, 10 kV/cm and 15 kV/cm, which was significantly lower than that of A549 cells(48.31%, 26.8%, and 5.96%, respectively for 8 kV/cm, 10 and 15 kV/cm). The induced apoptosis and necrosis rates were also significantly higher than A549(P<0.05); In addition, when exposed to 80 pulses(200 ns, 15 kV/cm), the ablation area of single-layer cells for A549/R was 1.59 times that of A549 cells, and the ablation threshold for A549/R(8 kV/cm) is significantly lower than that of A549(13 kV/cm). Therefore, the nanosecond pulsed electric field with a pulse width of 200 ns could preferentially kill A549/R, inducing a higher rate of apoptosis, ablation area and a lower ablation threshold. The experimental results showed that high-voltage nanosecond pulse could preferentially act on chemotherapy-resistant tumor cells.
Tumor seriously endanger the lives and health of human people. Chemoresistance is one of the main barriers to successful tumor treatment. In order to explore whether high voltage nanosecond pulsed electric fields can be equally sensitive to chemotherapy-resistant tumor cells compared with its homology tumor cells, lung cancer cell line A549 and cisplatin chemotherapy-resistant A549/R were used as the research object in this paper. After exposing to the 80 nanosecond pulses with a pulse width of 200 ns and a field strength of 5-15 kV/cm, the activity of cell, apoptosis and necrosis and ablation effect were analyzed and compared in groups. The results showed that the survival rate of A549/R was 5.65%, 7.78% and 2.80% respectively when exposed to pulses electric field with strength of 8 kV/cm, 10 kV/cm and 15 kV/cm, which was significantly lower than that of A549 cells(48.31%, 26.8%, and 5.96%, respectively for 8 kV/cm, 10 and 15 kV/cm). The induced apoptosis and necrosis rates were also significantly higher than A549(P<0.05); In addition, when exposed to 80 pulses(200 ns, 15 kV/cm), the ablation area of single-layer cells for A549/R was 1.59 times that of A549 cells, and the ablation threshold for A549/R(8 kV/cm) is significantly lower than that of A549(13 kV/cm). Therefore, the nanosecond pulsed electric field with a pulse width of 200 ns could preferentially kill A549/R, inducing a higher rate of apoptosis, ablation area and a lower ablation threshold. The experimental results showed that high-voltage nanosecond pulse could preferentially act on chemotherapy-resistant tumor cells.
引文
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[19]Ivey JW,Latouche EL,Sano MB,et al.Targeted cellular ablation based on the morphology of malignant cells[J].Scientific Reports,2015,5(1):17157.
[2]Devesa SS,Bray F,Vizcaino AP,et al.International lung cancer trends by histologic type:Male:Female differences diminishing and adenocarcinoma rates rising[J].International Journal of Cancer,2010,117(2):294-299.
[3]Mok TS,Wu YL,Thongprasert S,et al.Gefitinib or carboplatin-paclitaxel in pulmonary adenocarcinoma[J].The Journal of Evidence-Based Medicine,2009,361(10):947-957.
[4]Zeng-Rong N,Paterson J,Alpert L,et al.Elevated DNA repair capacity is associated with intrinsic resistance of lung cancer to chemotherapy[J].Cancer Research,1995,55(21):4760-4764.
[5]Martin LP,Hamilton TC,Schilder RJ.Platinum resistance:The role of DNA repair pathways[J].Clinical Cancer Research,2008,14(5):1291-1295.
[6]Townsend DM,Tew KD.The role of glutathione-S-transferase in anti-cancer drug resistance[J].Oncogene,2003,22(47):7369-7375.
[7]Oza AM.Clinical development of P glycoprotein modulators in oncology[J].Novartis Foundation Symposium,2002,243(1):103-115.
[8]Solyanik G.Multifactorial nature of tumor drug resistance[J].Experimental Oncology,2010,32(3):181-185.
[9]Zhang R,Yao W,Qian P,et al.Increased sensitivity of human lung adenocarcinoma cells to cisplatin associated with downregulated contactin-1[J].Biomedicine&Pharmacotherapy,2015,71(1):172-184.
[10]Miklav■i■ D,Ser■a G,Brecelj E,et al.Electrochemotherapy:technological advancements for efficient electroporation-based treatment of internal tumors[J].Medical&Biological Engineering&Computing,2012,50(12):1213-1225.
[11]Beebe SJ,Sain NM,Ren W.Induction of cell death mechanisms and apoptosis by nanosecond pulsed electric fields(nsPEFs)[J].Cells,2013,2(1):136-162.
[12]Schoenbach KH,Beebe SJ,Buescher ES.Intracellular effect of ultrashort electrical pulses[J].Bioelectromagnetics,2001,22(6):440-448.
[13]Schoenbach KH,Joshi RP,Kolb JF,et al.Ultrashort electrical pulses open a new gateway into biological cells[J].Proceedings of the IEEE,2004,92(7):1122-1137.
[14]姚陈果,孙才新,米彦,等.陡脉冲不可逆性电击穿治疗肿瘤的研究[J].高电压技术,2007,33(2):7-13.
[15]Nuccitelli R,Huynh J,Lui K,et al.Nanoelectroablation of human pancreatic carcinoma in a murine xenograft model without recurrence[J].International Journal of Cancer,2013,132(8):1933-1939.
[16]Nuccitelli R,Berridge JC,Mallon Z,et al.Nanoelectroablation of murine tumors triggers a CD8-dependent inhibition of secondary tumor growth[J].PLoS ONE,2015,10(7):e0134364.
[17]Nuccitelli R,Mcdaniel A,Anand S,et al.Nano-pulse stimulation is a physical modality that can trigger immunogenic tumor cell death[J].Journal for Immunotherapy of Cancer,2017,5(1):32.
[18]Yao C,Hu X,Mi Y,et al.Window effect of pulsed electric field on biological cells[J].IEEE Transactions on Dielectrics&Electrical Insulation,2009,16(5):1259-1266.
[19]Ivey JW,Latouche EL,Sano MB,et al.Targeted cellular ablation based on the morphology of malignant cells[J].Scientific Reports,2015,5(1):17157.
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