Full Text:   <1236>

Summary:  <468>

CLC number: R735; R457

On-line Access: 2015-05-04

Received: 2014-08-11

Revision Accepted: 2015-01-22

Crosschecked: 2015-04-10

Cited: 2

Clicked: 2384

Citations:  Bibtex RefMan EndNote GB/T7714

 ORCID:

Jin-ting Yang

http://orcid.org/0000-0003-3859-6340

-   Go to

Article info.
Open peer comments

Journal of Zhejiang University SCIENCE B 2015 Vol.16 No.5 P.395-403

http://doi.org/10.1631/jzus.B1400224


Cisplatin combined with hyperthermia kills HepG2 cells in intraoperative blood salvage but preserves the function of erythrocytes


Author(s):  Jin-ting Yang, Li-hui Tang, Yun-qing Liu, Yin Wang, Lie-ju Wang, Feng-jiang Zhang, Min Yan

Affiliation(s):  Department of Anesthesiology, the Second Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou 310009, China; more

Corresponding email(s):   zryanmin@zju.edu.cn

Key Words:  Erythrocytes, HepG2 cells, Intraoperative blood salvage, Cisplatin, Hyperthermia


Jin-ting Yang, Li-hui Tang, Yun-qing Liu, Yin Wang, Lie-ju Wang, Feng-jiang Zhang, Min Yan. Cisplatin combined with hyperthermia kills HepG2 cells in intraoperative blood salvage but preserves the function of erythrocytes[J]. Journal of Zhejiang University Science B, 2015, 16(5): 395-403.

@article{title="Cisplatin combined with hyperthermia kills HepG2 cells in intraoperative blood salvage but preserves the function of erythrocytes",
author="Jin-ting Yang, Li-hui Tang, Yun-qing Liu, Yin Wang, Lie-ju Wang, Feng-jiang Zhang, Min Yan",
journal="Journal of Zhejiang University Science B",
volume="16",
number="5",
pages="395-403",
year="2015",
publisher="Zhejiang University Press & Springer",
doi="10.1631/jzus.B1400224"
}

%0 Journal Article
%T Cisplatin combined with hyperthermia kills HepG2 cells in intraoperative blood salvage but preserves the function of erythrocytes
%A Jin-ting Yang
%A Li-hui Tang
%A Yun-qing Liu
%A Yin Wang
%A Lie-ju Wang
%A Feng-jiang Zhang
%A Min Yan
%J Journal of Zhejiang University SCIENCE B
%V 16
%N 5
%P 395-403
%@ 1673-1581
%D 2015
%I Zhejiang University Press & Springer
%DOI 10.1631/jzus.B1400224

TY - JOUR
T1 - Cisplatin combined with hyperthermia kills HepG2 cells in intraoperative blood salvage but preserves the function of erythrocytes
A1 - Jin-ting Yang
A1 - Li-hui Tang
A1 - Yun-qing Liu
A1 - Yin Wang
A1 - Lie-ju Wang
A1 - Feng-jiang Zhang
A1 - Min Yan
J0 - Journal of Zhejiang University Science B
VL - 16
IS - 5
SP - 395
EP - 403
%@ 1673-1581
Y1 - 2015
PB - Zhejiang University Press & Springer
ER -
DOI - 10.1631/jzus.B1400224


Abstract: 
The safe use of intraoperative blood salvage (IBS) in cancer surgery remains controversial. Here, we investigated the killing effect of cisplatin combined with hyperthermia on human hepatocarcinoma (HepG2) cells and erythrocytes from IBS in vitro. hepG2 cells were mixed with concentrated erythrocytes and pretreated with cisplatin (50, 100, and 200 μg/ml) alone at 37 °C for 60 min and cisplatin (25, 50, 100, and 200 μg/ml) combined with hyperthermia at 42 °C for 60 min. After pretreatment, the cell viability, colony formation and DNA metabolism in HepG2 and the Na+-K+-ATPase activity, 2,3-diphosphoglycerate (2,3-DPG) concentration, free hemoglobin (Hb) level, osmotic fragility, membrane phosphatidylserine externalization, and blood gas variables in erythrocytes were determined. Pretreatment with cisplatin (50, 100, and 200 μg/ml) combined with hyperthermia (42 °C) for 60 min significantly decreased HepG2 cell viability, and completely inhibited colony formation and DNA metabolism when the HepG2 cell concentration was 5×104 ml−1 in the erythrocyte (P<0.01). Erythrocytic Na+-K+-ATPase activity, 2,3-DPG level, phosphatidylserine externalization, and extra-erythrocytic free Hb were significantly altered by hyperthermia plus high concentrations of cisplatin (100 and 200 μg/ml) (P<0.05), but not by hyperthermia plus 50 μg/ml cisplatin (P>0.05). In conclusion, pretreatment with cisplatin (50 μg/ml) combined with hyperthermia (42 °C) for 60 min effectively eliminated hepG2 cells from IBS but did not significantly affect erythrocytes in vitro.

顺铂联合热处理对术中回收血红细胞及其中混杂的肝肿瘤细胞株(HepG2)的影响

目的:研究离体热处理和顺铂联合热处理对术中回收血红细胞功能的影响及其中混杂的肝肿瘤细胞株(HepG2)的杀灭作用。
创新点:(1)采用多种评价指标研究了不同时间离体热处理对术中回收血中混杂的HepG2的杀灭作用及对红细胞的影响,并确定了对红细胞安全且能有效杀灭HepG2的离体热处理时间。(2)从多个角度评价了离体顺铂联合热处理对术中回收血红细胞的影响及对其中混杂的HepG2的杀灭作用,确定了该方案中对红细胞安全且能有效去除HepG2的顺铂浓度。
方法:采用3-(4,5-二甲基噻唑-2)-2,5-二苯基四氮唑溴盐(MTT)、5-乙炔基-2’脱氧尿嘧啶核苷(EdU)和平板克隆形成评估HepG2的细胞存活率、DNA复制率和克隆形成能力(图1和3);从红细胞渗透脆性、携氧能力(2,3-二磷酸甘油酸(2,3-DPG)、半饱和氧分压(P50))、能量代谢(Na+-K+-ATPase、pH)、膜完整性(游离血红蛋白(Hb)、血清K+和Na+浓度、细胞膜磷脂酰丝氨酸外翻比例)等角度评估红细胞功能(图2和4;表3)。
结论:肝肿瘤术中回收血经离体顺铂联合热处理(42°C,50µg/ml)60min后,能有效清除其中混杂的HepG2,但对红细胞无显著影响,值得体内进一步研究顺铂热处理有效应用于肿瘤手术自体血液回输的安全方案。

关键词:红细胞;HepG2细胞株;术中血液回收;顺铂;热处理

Darkslateblue:Affiliate; Royal Blue:Author; Turquoise:Article

Reference

[1]Adias, T.C., Jeremiah, Z., Uko, E., et al., 2006. Autologous blood transfusion—a review. S. Afr. J. Surg., 44(3):114-116, 118.

[2]Catling, S., Williams, S., Freites, O., et al., 2008. Use of a leucocyte filter to remove tumour cells from intra-operative cell salvage blood. Anaesthesia, 63(12):1332-1338.

[3]Edelman, M.J., Potter, P., Mahaffey, K.G., et al., 1996. The potential for reintroduction of tumor cells during intraoperative blood salvage: reduction of risk with use of the RC-400 leukocyte depletion filter. Urology, 47(2):179-181.

[4]Fӧller, M., Braun, M., Qadri, S.M., et al., 2010. Temperature sensitivity of suicidal erythrocyte death. Eur. J. Clin. Invest., 40(6):534-540.

[5]Hansen, E., Wolff, N., Knuechel, R., et al., 1995. Tumor cells in blood shed from the surgical field. Arch. Surg., 130(4): 387-393.

[6]Hansen, E., Knuechel, R., Altmeppen, J., et al., 1999. Blood irradiation for intraoperative autotransfusion in cancer surgery: demonstration of efficient elimination of contaminating tumor cells. Transfusion, 39(6):608-615.

[7]Harrison, L.E., Bryan, M., Pliner, L., et al., 2008. Phase I trial of pegylated liposomal doxorubicin with hyperthermic intraperitoneal chemotherapy in patients undergoing cytoreduction for advanced intra-abdominal malignancy. Ann. Surg. Oncol., 15(5):1407-1413.

[8]Hartmann, S., Kriegebaum, U., Küchler, N., et al., 2014. Correlation of MAGE-A tumor antigens and the efficacy of various chemotherapeutic agents in head and neck carcinoma cells. Clin. Oral Investig., 18(1):189-197.

[9]Hasegawa, A., Shimizu, R., Mohandas, N., et al., 2012. Mature erythrocyte membrane homeostasis is compromised by loss of the GATA1-FOG1 interaction. Blood, 119(11):2615-2623.

[10]Herron, D.M., Grabowy, R., Connolly, R., et al., 1997. The limits of bloodwarming: maximally heating blood with an inline microwave bloodwarmer. J. Trauma, 43(2):219-228.

[11]Hildebrandt, B., Wust, P., Ahlers, O., et al., 2002. The cellular and molecular basis of hyperthermia. Crit. Rev. Oncol. Hematol., 43(1):33-56.

[12]Issels, R.D., 2008. Hyperthermia adds to chemotherapy. Eur. J. Cancer, 44(17):2546-2554.

[13]Jemal, A., Center, M.M., Desantis, C., et al., 2010. Global patterns of cancer incidence and mortality rates and trends. Cancer Epidemiol. Biomarkers Prev., 19(8):1893-1907.

[14]Kiefer, C.R., Snyder, L.M., 2000. Oxidation and erythrocyte senescence. Curr. Opin. Hematol., 7(2):113-116.

[15]Kumar, N., Chen, Y., Zaw, A.S., et al., 2014. Use of intraoperative cell-salvage for autologous blood transfusions in metastatic spine tumour surgery: a systematic review. Lancet Oncol., 15(1):e33-e41.

[16]Moretto, J., Chauffert, B., Ghiringhelli, F., et al., 2011. Discrepancy between in vitro and in vivo antitumor effect of a new platinum(II) metallointercalator. Invest. New Drugs, 29(6):1164-1176.

[17]Sachar, S., Saxena, R.K., 2011. Cytotoxic effect of poly-dispersed single walled carbon nanotubes on erythrocytes in vitro and in vivo. PLoS ONE, 6(7):e22032.

[18]Su, X.Y., Yin, H.T., Li, S.Y., et al., 2012. Intervention effects of nedaplatin and cisplatin on proliferation and apoptosis of human tumour cells in vitro. Asian Pac. J. Cancer Prev., 13(9):4531-4536.

[19]Sugarbaker, P.H., 2007. Laboratory and clinical basis for hyperthermia as a component of intracavitary chemotherapy. Int. J. Hyperthermia, 23(5):431-442.

[20]Wang, X., Ji, B., Zhang, Y., et al., 2012. Comparison of the effects of three cell saver devices on erythrocyte function during cardiopulmonary bypass procedure—a pilot study. Artif. Organs, 36(10):931-935.

[21]Waters, J.H., Donnenberg, A.D., 2009. Blood salvage and cancer surgery: should we do it? Transfusion, 49(10):2016-2018.

[22]Waters, J.H., Yazer, M., Chen, Y.F., et al., 2012. Blood salvage and cancer surgery: a meta-analysis of available studies. Transfusion, 52(10):2167-2173.

[23]Westermann, A.M., Grosen, E.A., Katschinski, D.M., et al., 2001. A pilot study of whole body hyperthermia and carboplatin in platinum-resistant ovarian cancer. Eur. J. Cancer, 37(9):1111-1117.

[24]Winslow, R.M., 2007. The role of hemoglobin oxygen affinity in oxygen transport at high altitude. Respir. Physiol. Neurobiol., 158(2-3):121-127.

[25]Wu, J.B., Lei, W.F., Zhao, L.Y., et al., 2006. Hyperthermia: its effectiveness in killing tumor cells and influence on Na+-K+-ATPase activities in erythrocytes. Chin. J. Anesthesiol., 26(3):258-260 (in Chinese).

[26]Zhou, J.F., Wang, P., Lei, W.F., 2011. Effectiveness of cis-diamminedichloroplatinum combined with hyperthermia in killing liver tumor cells and its influence on erythrocytes in vitro. Chin. J. Anesthesiol., 31(2):193-196 (in Chinese).

Open peer comments: Debate/Discuss/Question/Opinion

<1>

Please provide your name, email address and a comment





Journal of Zhejiang University-SCIENCE, 38 Zheda Road, Hangzhou 310027, China
Tel: +86-571-87952783; E-mail: cjzhang@zju.edu.cn
Copyright © 2000 - Journal of Zhejiang University-SCIENCE