Low-frequency ultrasound with ultrasound contrast agent reverses paclitaxel resistance in prostate cancer cells by activating endoplasmic reticulum stress

Yuqi WU; Xiangwei WANG

doi:10.16016/j.1000-5404.201711190

您当前的位置：

首页 >

文章列表页 >

Low-frequency ultrasound with ultrasound contrast agent reverses paclitaxel resistance in prostate cancer cells by activating endoplasmic reticulum stress

Basic Medicine | 更新时间：2026-01-09

- Low-frequency ultrasound with ultrasound contrast agent reverses paclitaxel resistance in prostate cancer cells by activating endoplasmic reticulum stress
- Journal of Third Military Medical University Vol. 40, Issue 8, Pages: 673-678(2018)
- 作者机构：
  
  400037 重庆，陆军军医大学(第三军医大学)第二附属医院泌尿外科
- 作者简介：
- 基金信息：
  
  the General Program of National Natural Science Foundation of China(81172443)
- DOI：10.16016/j.1000-5404.201711190
  CLC： R445.1;R737.25;R979.1
- Received：22 November 2017，
  
  Revised：2017-12-19，
  
  Published：30 April 2018
- 稿件说明：
移动端阅览
Yuqi WU, Xiangwei WANG. Low-frequency ultrasound with ultrasound contrast agent reverses paclitaxel resistance in prostate cancer cells by activating endoplasmic reticulum stress[J]. Journal of Third Military Medical University, 2018, 40(8): 673-678.
DOI：

Yuqi WU, Xiangwei WANG. Low-frequency ultrasound with ultrasound contrast agent reverses paclitaxel resistance in prostate cancer cells by activating endoplasmic reticulum stress[J]. Journal of Third Military Medical University, 2018, 40(8): 673-678. DOI： 10.16016/j.1000-5404.201711190.

摘要

目的

探讨低频超声联合微泡造影剂辅助紫杉醇逆转前列腺癌耐药的作用和机制。

方法

筛选超声辐照紫杉醇耐药前列腺癌细胞株PC-3R的非毒性照射时间；将PC-3R细胞随机分为对照组、紫杉醇组(PTX组)、低频超声组(LFUS组)、紫杉醇和低频超声组(PTX+LFUS组)，以及紫杉醇和低频超声联合微泡剂组(PTX+LFUS+UCA组)，分别进行干预。AnnexinV-FITC/PI双染法检测各组细胞凋亡；q-PCR法检测各组细胞Bcl-2和多耐药基因表达变化；Western blot检测各组细胞GRP78和c-jun蛋白表达。GRP78 siRNA转染PC-3R细胞，微泡和低频超声联合处理后，Western blot检测GRP78蛋白表达，Annexin V-FITC/PI双染法检测凋亡情况。

结果

低频超声照射频率1 MHz，强度1.2 W/cm

，非毒性照射时间为10 s。与单纯应用紫杉醇相比低频超声辅助紫杉醇处理前列腺耐药细胞可显著提高细胞凋亡率(

0.01)，联合微泡剂照射后，凋亡率进一步提高(

0.01)，并下调Bcl-2和多耐药基因的表达。PTX+LFUS+UCA组内质网应激相关蛋白GRP78的表达较PTX组和PTX+LFUS组升高，c-jun的表达下降(

0.01)。应用siRNA干扰GRP78基因表达后，细胞凋亡率下降(

0.01)。

结论

低频超声联合微泡造影剂通过激活内质网应激，抑制其下游JNK/c-jun通路下调多耐药基因及凋亡抑制基因的表达，提高化疗药物诱导细胞凋亡的作用，逆转肿瘤细胞的耐药性。

Abstract

Objective

To investigate the effect of low-frequency ultrasound (LFUS) combined with ultrasound contrast agent (UCA) on reversing paclitaxel (PTX) resistance in prostate cancer cells and explore the mechanism.

Methods

The non-toxic ultrasound irradiation time was determined in PTX-resistant prostate cancer PC-3 (PC-3R) cells. The cells were then divided into control group

paclitaxel (PTX) group

LFUS group

PTX+LFUS group

and PTX+LFUS+UCA group for corresponding treatments. After the treatments

the apoptotic rates of PC-3R cells were determined using flow cytometry

and the protein levels of GRP78 and c-jun and the mRNA levels of Bcl-2 and multidrug resistance-associated proteins (MRPs) were detected using Western blotting and q-PCR

respectively. A lentiviral vector expressing siRNA targeting human GRP78 was transfected in the cells prior to exposure to LFUS with UCA

and the protein expression level of GRP78 was determined by Western blotting and cell apoptosis was analyzed using flow cytometry.

Results

Under the optimal settings (1 MHz

1.2 W/cm

50% duty cycle

and 10 s)

LFUS exposu

re combined with PTX treatment significantly increased the apoptotic rate of prostate cancer cells compared with PTX treatment alone (

0.01); in the presence of UCA

LFUS plus PTX treatment further increased the cell apoptosis rate (

0.01) and down-regulated the expressions of Bcl-2 and MRPs. LFUS with UCA plus PTX treatment resulted in significantly increased expression of endoplasmic reticulum stress-related protein GRP78 and lowered c-jun expression as compared LFUS plus PTX treatment without UCA. Transfection of the cells with siRNA targeting GRP78 caused a significant decrease of the cell apoptosis rate following LFUS with UCA plus PTX treatment (

0.01).

Conclusion

LFUS combined with UCA can inhibit the expression of MRPs and the apoptosis-suppressing gene by down-regulating the JNK/c-jun pathway in the endoplasmic reticulum to enhance the effect of PTX-induced apoptosis and reverse drug resistance of prostate cancer cells.

关键词

Keywords

references

SIEGEL R, MA J, ZOU Z, et al . Cancer statistics, 2014[J ] . CA Cancer J Clin, 2014, 64(1): 9-29. DOI: 10.3322/caac.21208.

LEE S, KIM K, HO JN, et al . Analysis of resistance-associated gene expression in docetaxel-resistant prostate cancer cells[J ] .Oncol Lett, 2017, 14(3): 3011-3018. DOI: 10.3892/ol.2017.6541.

SHEN Z Y, XIA G L, WU M F, et al . The effects of low-frequency ultrasound and microbubbles on rabbit hepatic tumors[J ] . ExpBiol Med (Maywood), 2014, 239(6): 747-757. DOI: 10.1177/1535370214525320.

WAN G Y, LIU Y, CHEN B W, et al . Recent advances of sonodynamic therapy in cancer treatment[J ] . Cancer Biol Med, 2016, 13(3): 325-338.DOI: 10.20892/j.issn.2095-3941.2016.0068.

MCHALE A P, CALLAN J F, NOMIKOU N, et al . Sono-dynamic therapy: concept, mechanism and application to cancer treatment[J ] . Adv Exp Med Biol, 2016, 880: 429-450. DOI: 10.1007/978-3-319-22536-4_22.

PITT W G, HUSSEINI G A, ROEDER B L, et al . Preliminary results of combining low frequency low intensity ultrasound and liposomal drug delivery to treat tumors in rats[J ] . J Nanosci Nanotechnol, 2011, 1 1(3): 1866-1870. DOI: 10.1166/jnn.2011.3117.

HOU R, XU Y, LU Q, et al . Effect of low-frequency low-intensity ultrasound with microbubbles on prostate cancer hypoxia[J ] . Tumour Biol, 2017, 39(10): 1-9. DOI: 10.1177/1010428317719275.

朱燕, 刘政, 杨鹰, 等.低频超声联合微泡增强宫颈癌Hela细胞对顺铂敏感性的实验研究[J].第三军医大学学报, 2016, 38(15): 1741-1745. DOI: 10.16016/j.1000-5404.201511184.

ZHU Y, LIU Z, YANG Y, et al . Low-frequency ultrasound combined with microbubble agent enhances cytotoxicity of cisplatin to Hela cells[J ] . J Third Mil Med Univ, 2016, 38(15): 1741-1745. DOI: 10.16016/j.1000-5404.201511184.

SNIPSTAD S, BERG S, MORCH Y, et al . Ultrasound Improves the Delivery and Therapeutic Effect of Nanoparticle-Stabilized Microbubbles in Breast Cancer Xenografts[J ] . Ultrasound Med Biol, 2017, 43(11): 2651-2669. DOI: 10.1016/j.ultrasmedbio.2017.06.029.

HU J L, HU X L, GUO A Y, et al . Endoplasmic reticulum stress promotes autophagy and apoptosis and reverses chemoresistance in human ovarian cancer cells[J ] . Oncotarget, 2017, 8(30): 49380-49394. DOI: 10.18632/oncotarget.17673.

OGAWA H, KAIRA K, TAKAHASHI K, et al . Prognostic role of BiP/GRP78 expression as ER stress in patients with gastric adenocarcinoma[J ] . Cancer Biomark, 2017, 20(3): 273-281. DOI: 10.3233/CBM-170062.

CAI J, DU S, WANG H, et al . Tenascin-C induces migration and invasion through JNK/c-Jun signalling in pancreatic cancer[J ] . Oncotarget, 2017, 8(43): 74406.

XIONG Z, JIANG R, LI X, et al . Different roles of GRP78 on cell proliferation and apoptosis in cartilage development[J ] . Int J MolSci, 2015, 16(9): 21153-21176. DOI: 10.3390/ijms160921153.

POZNIAK B, PAWLAK A, OBMINSKA-MRUKOWICA B. Flow cytometric assessment of P-glycoprotein and multidrug resistance-associated protein activity and expression in canine lymphoma[J ] . In Vivo , 2015, 29(1): 149-153.

SUI H, CAI G X, PAN S F, et al . miR200c attenuates P-gp-mediated MDR and metastasis by targeting JNK2/c-Jun signaling pathway in colorectal cancer[J ] . Mol Cancer Ther, 2014, 13(12): 3137-3151. DOI: 10.1158/1535-7163.MCT-14-0167.

朱江, 陈青, 田思, 等.光敏剂MPPa介导光动力诱导人乳腺癌细胞MCF-7的内质网应激途径凋亡[J].第三军医大学学报, 2015, 37(21): 2115-2120. DOI: 10.16016/j.1000-5404.201503144.

ZHU J, CHEN Q, TIAN S, et al . Pyropheophorbide-a methyl ester-mediated photodynamic therapy induces apoptosis in human breast cancer MCF-7cells through endoplasmic reticulum stress pathway[J ] .J Third Mil Med Univ, 2015, 37(21): 2115-2120. DOI: 10.16016/j.1000-5404.201503144.

Views

下载量

CSCD

Alert me when the article has been cited

提交

Tools

Publicity Resources

Preparation of nanosized hypoxia-inducible factor 1α inhibitor and its application in reversing chemoresistance in colorectal cancer cells

Prostate cancer T-stage intelligent diagnosis based on MRI images and deep learning

Risk factors for biochemical recurrence of prostate cancer after radical prostatectomy

Dihydroartemisinin induces pyroptosis and regulates apoptosis-associated speck-like protein methylation in prostate cancer cells

Silencing high mobility group protein B1 inhibits malignant biological behavior of glioblastoma cells

Related Author

ZHENG Chuanhao

DAN Ruijue

XI Xi

YING Wang

LUO Qiang

许杉杉

呼延若曦

吴哲

Related Institution

Department of Gastroenterology， Second Affiliated Hospital， Army Medical University （Third Military Medical University）

Department of Pharmacognosy, Faculty of Pharmacy and Laboratory Medicine, Army Military Medical University (Third Military Medical University)

Center of Hematology, Second Affiliated Hospital, Army Medical University(Third Military Medical University)

Institute of Pathology, First Affiliated Hospital, Army Medical University(Third Military Medical University)

⁰