納秒脈沖電場(chǎng)消融乳腺癌的實(shí)驗(yàn)研究

謝貴林++++++陳璐艷++++++陳新華+++++溫浩++++++陳新梅

[摘要] 目的 探索納秒脈沖對(duì)乳腺癌高轉(zhuǎn)移細(xì)胞株MDA-MB-231的治療效果及其分子機(jī)制。 方法 納秒脈沖電消融乳腺癌細(xì)胞及荷瘤小鼠后，檢測(cè)接受納秒脈沖治療（實(shí)驗(yàn)組）及未接受納秒脈沖治療（對(duì)照組）的小鼠腫瘤細(xì)胞增殖、周期和凋亡相關(guān)指標(biāo)，觀察腫瘤的體積、生長(zhǎng)形態(tài)及速度等的變化。 結(jié)果 納秒脈沖在體外可以顯著降低細(xì)胞活性，誘導(dǎo)細(xì)胞凋亡，表現(xiàn)為實(shí)驗(yàn)組CCK-8細(xì)胞活性抑制率與對(duì)照組比較，差異有高度統(tǒng)計(jì)學(xué)意義（P < 0.01），且各實(shí)驗(yàn)組間差異均有高度統(tǒng)計(jì)學(xué)意義（P < 0.01）；投射電鏡和線粒體電勢(shì)實(shí)驗(yàn)觀察到細(xì)胞出現(xiàn)凋亡，抑制腫瘤生長(zhǎng)。 結(jié)論 納秒脈沖可以通過誘導(dǎo)乳腺癌細(xì)胞MDA-MB-231的凋亡而對(duì)腫瘤進(jìn)行消融。

[關(guān)鍵詞] 乳腺癌；MDA-MB-231；納秒脈沖；消融

[中圖分類號(hào)] R730.5 [文獻(xiàn)標(biāo)識(shí)碼] A [文章編號(hào)] 1673-7210（2015）06（a）-0004-06

Experiment study of nanosecond pulsed electric fields ablate breast cancer

XIE Guilin1 CHEN Luyan2 CHEN Xinhua2 WEN Hao3 CHEN Xinmei4

1.Department of General Surgery， the Second Hospital of Shaoxing， Zhejiang Province， Shaoxing 312000， China； 2.Department of General Surgery， the First Affiliated Hospital of Zhejiang University， Zhejiang Province， Hangzhou 310000， China； 3.Department of General Surgery， the First Affiliated Hospital of Xinjiang Medical University， Xinjiang Uygur Autonomous Region， Urumqi 830054， China； 4. School of Pharmacy， Shandong University of Traditional Chinese Medicine， Shandong Province， Ji'nan 250355， China

[Abstract] Objective To investigate the treatment effect of nanosecond pulsed electric fields （nsPEFs） on highly-metastatic human breast cancer cell line MDA-MB-231 in vitro and in vivo the and the possible molecular mechanism. Methods After treatment of nsPEFs in different parameters， the apoptosis， proliferation and invasion of MDA-MB-231 cells in rats of nanosecond pulse therapy （experimental group） and non-nanosecond pulse therapy （control group） were examined with CCK-8， TUNEL， flow cytometer， transmission electron microscopy （TEM）. The volum of tumor size， groth pattern and speed were observed. Results In vitro， nsPEFs inhibited the cell viability and induced cell apoptosis. The cell viability of CCK-8 in the experimental group was reduced significantly compared with control group， the difference was statistically significant （P < 0.01）， and the differences among different experimental group were statistically significant （P < 0.01）； it also presented mitochondria degeneration. Flow cytometer showed that nsPEFs could inhibit cancer cell proliferation. Conclusion nsPEFs can ablate human breast cancer MDA-MB-231 by activating caspase signaling pathway.

[Key words] Breast cancer； MDA-MB-231； Nanosecond pulsed electric fields （nsPEFs）； Ablation

乳腺癌成為威脅女性的重大健康問題[1]。根據(jù)我國(guó)2014年腫瘤年報(bào)，乳腺癌發(fā)病率和病死率持續(xù)增高[2]。MDA-MB-231可以用于制作研究高侵襲乳癌細(xì)胞的細(xì)胞模型和動(dòng)物模型[3]。目前多學(xué)科聯(lián)合治療是乳腺癌的主要方法[4]。根據(jù)目前消融介入技術(shù)的發(fā)展，微創(chuàng)消融也逐漸進(jìn)入乳腺癌的綜合治療計(jì)劃中。脈沖場(chǎng)強(qiáng)可以引起電穿孔[5]，電穿孔不僅提高細(xì)胞膜對(duì)化療藥物的滲透率，增強(qiáng)化療藥物細(xì)胞毒性，還能通過降低化療藥物的使用劑量從而減小對(duì)正常細(xì)胞的毒副作用。而納秒脈沖是不同于電穿孔技術(shù)的最新一代電脈沖消融技術(shù)。納秒脈沖引起細(xì)胞膜的微孔出現(xiàn)，最終形成細(xì)胞凋亡[6-7]。避免了嚴(yán)重的炎性反應(yīng)[8]，凋亡最初是在胚胎[9]及免疫系統(tǒng)[10]發(fā)育中發(fā)現(xiàn)。而在腫瘤治療中凋亡產(chǎn)生則意味著腫瘤被消滅而機(jī)體未被傷害。Caspase是凋亡發(fā)生的一個(gè)重要分子標(biāo)志[11-13]，在本實(shí)驗(yàn)中納秒級(jí)脈單獨(dú)治療腫瘤，避免了化療副作用。