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低剂量重离子辐照对胰腺癌患者外周血淋巴细胞亚群百分比及细胞因子表达的影响

[Effect of low dose heavy ion irradiation on subset percentage and cytokines expression of peripheral blood lymphocytes in patients with pancreatic cancer].

作者信息

Chen Yingtai, Wang Chenfeng, He Mingyan, Zhang Hong, Chen Xuezhong

机构信息

Department of Abdominal Surgical Oncology, Cancer Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100021, China.

Email:

出版信息

Zhonghua Zhong Liu Za Zhi. 2014 Jun;36(6):435-9.

Abstract

OBJECTIVE

The aim of this study was to examine the effect of low dose heavy ion irradiation on the subset percentage and expression of cytokines of peripheral blood lymphocytes(PBL) in patients with pancreatic cancer.

METHODS

PBL from 21 patients with pancreatic cancer were divided into three groups: sham, X-ray and ¹²C⁶⁺ irradiation groups, and the cell responses were measured at 24 hours after radiation exposure. The percentages of T and NK cell subsets were detected by flow cytometry. The mRNA expression of interleukin (IL)-2, tumor necrosis factor (TNF)-α and interferon (IFN)-γ were examined by real-time quantitative RT-PCR (qRT-PCR). The cytokine protein levels in supernatant of cultured cells were assayed by enzyme-linked immunosorbent assays (ELISA).

RESULTS

The percentage of T lymphocyte subsets was significantly increased at 24 hours after exposure to low dose radiation, and the effect was more pronounced in the group receiving 0.05 Gy ¹²C⁶⁺ ion irradiation than that in the group receiving X-ray irradiation [CD3⁺ T cells: (67.15 ± 4.36)% vs. (60.81 ± 8.35)%; CD3⁺ CD4⁺ T cells: (19.02 ± 2.35)% vs. (17.21 ± 2.86)%; CD3⁺ CD8⁺ T cells: (46.59 ± 6.07)% vs. (41.18 ± 6.35)%. (P < 0.05 for all)]. However, there were no significant changes in the CD3⁺ CD4⁺/CD3⁺ CD8⁺ ratio (0.67 for sham, 0.65 for X-ray, and 0.68 for ¹²C⁶⁺ groups) and percentage of NK cell subsets (P > 0.05 for all). Expression levels of IFN-γ mRNA (cycle threshold/CT value was 23.35 ± 3.16 for ¹²C⁶⁺, CT value was 27.25 ± 2.15 for X-ray) and IL-2 (CT value was 24.19 ± 3.56 for ¹²C⁶⁺, CT value was 27.85 ± 4.08 for X-ray) in PBL, and their protein levels in the supernatant were significantly increased at 24 hours after exposure to the low dose radiation (P < 0.05). The effects were more pronounced in the group receiving 0.05 Gy ¹²C⁶⁺ ion irradiation than that in the group receiving X-ray irradiation. However, there was no significant change in the TNF-α production of PBL.

CONCLUSIONS

Low dose irradiation may alleviate the immune suppression caused by tumor burden and that the effect is more pronounced for 0.05 Gy high linear energy transfer (LET) ¹²C⁶⁺ irradiation. The percentage of T cell subsets and cytokines production could be used as sensitive indicators of acute response to low dose irradiation.

摘要

目的

本研究旨在探讨低剂量重离子辐射对胰腺癌患者外周血淋巴细胞(PBL)亚群百分比及细胞因子表达的影响。

方法

将21例胰腺癌患者的PBL分为三组:假照射组、X射线照射组和¹²C⁶⁺照射组,并在辐射暴露后24小时测量细胞反应。通过流式细胞术检测T和NK细胞亚群的百分比。采用实时定量逆转录聚合酶链反应(qRT-PCR)检测白细胞介素(IL)-2、肿瘤坏死因子(TNF)-α和干扰素(IFN)-γ的mRNA表达。通过酶联免疫吸附测定(ELISA)检测培养细胞上清液中的细胞因子蛋白水平。

结果

低剂量辐射暴露后24小时,T淋巴细胞亚群百分比显著增加,且接受0.05 Gy ¹²C⁶⁺离子照射组的效果比接受X射线照射组更明显[CD3⁺ T细胞:(67.15 ± 4.36)%对(60.81 ± 8.35)%;CD3⁺ CD4⁺ T细胞:(19.02 ± 2.35)%对(17.21 ± 2.86)%;CD3⁺ CD8⁺ T细胞:(46.59 ± 6.07)%对(41.18 ± 6.35)%。所有P < 0.05]。然而,CD3⁺ CD4⁺/CD3⁺ CD8⁺比值(假照射组为0.67,X射线照射组为0.65,¹²C⁶⁺照射组为0.68)和NK细胞亚群百分比无显著变化(所有P > 0.05)。PBL中IFN-γ mRNA表达水平(¹²C⁶⁺照射组的循环阈值/CT值为23.35 ± 3.16,X射线照射组的CT值为27.25 ± 2.15)和IL-2(¹²C⁶⁺照射组的CT值为24.19 ± 3.56,X射线照射组的CT值为27.85 ± 4.08)及其上清液中的蛋白水平在低剂量辐射暴露后24小时显著增加(P < 0.05)。接受0.05 Gy ¹²C⁶⁺离子照射组的效果比接受X射线照射组更明显。然而,PBL的TNF-α产生无显著变化。

结论

低剂量照射可能减轻肿瘤负荷引起的免疫抑制,且对于0.05 Gy高传能线密度(LET)的¹²C⁶⁺照射效果更明显。T细胞亚群百分比和细胞因子产生可作为低剂量照射急性反应的敏感指标。

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