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超声辐射及 SonoVue 微泡介导的 PRR11 靶向 RNA 干扰抑制乳腺癌细胞增殖和转移,但促进细胞凋亡。

Ultrasonic irradiation and SonoVue microbubbles-mediated RNA interference targeting PRR11 inhibits breast cancer cells proliferation and metastasis, but promotes apoptosis.

机构信息

Department of Ultrasound, Shenzhen People’s Hospital, The Second Clinical Medical College of Jinan University, Shenzhen, China.

Shenzhen Medical Ultrasound Engineering Center, Shenzhen People’s Hospital, The Second Clinical Medical College of Jinan University, Shenzhen, China.

出版信息

Biosci Rep. 2020 Nov 27;40(11). doi: 10.1042/BSR20201854.

Abstract

The present study compared the effects of ultrasonic irradiation and SonoVue microbubbles (US) or Lipofectamine 3000 on the transfection of small interfering RNA for PRR11 (siPRR11) and Proline-rich protein 11 (PRR11) overexpression plasmid into breast cancer cells. SiPRR11 and PRR11 overexpression plasmid were transfected into breast cancer MCF7 cells mediated by US and Lipofectamine 3000. PRR11 expressions in breast cancer and normal tissues were determined using Gene Expression Profiling Interactive Analysis (GEPIA). The viability, proliferation, migration, invasion and apoptosis of breast cancer cells were respectively measured by MTT assay, clone formation assay, scratch wound-healing assay, Transwell assay and flow cytometry. PRR11 and epithelial-to-mesenchymal transition (EMT)-related and apoptosis-related (B-cell lymphoma 2, Bcl-2; Bcl-2-associated protein X, Bax) proteins' expressions were detected by quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot as appropriate. As ultrasonic intensity increased, the viability of MCF7 cells was decreased. Results from GEPIA suggested that PRR11 was up-regulated in breast cancer. Silencing PRR11 mediated by US showed a higher efficiency than by Lipofectamine 3000. SiPRR11 transfected by Lipofectamine 3000 suppressed cells growth and metastasis, while promoted cell apoptosis. Moreover, E-cadherin (E-cad) and Bax expressions were high but N-cadherin (N-cad), Snail and Bcl-2 expressions were low. However, overexpressed PRR11 caused the opposite effects. More importantly, transfection of siPRR11 and PRR11 overexpression plasmid using US had a higher efficacy than using Lipofectamine 3000. US transfection of PRR11 siRNA showed better effects on inhibiting breast cancer progression. The current findings contribute to a novel treatment for breast cancer.

摘要

本研究比较了超声辐射和 SonoVue 微泡(US)或 Lipofectamine 3000 对 PRR11(siPRR11)和富含脯氨酸蛋白 11(PRR11)过表达质粒转染乳腺癌细胞的影响。通过 US 和 Lipofectamine 3000 将 siPRR11 和 PRR11 过表达质粒转染乳腺癌 MCF7 细胞。使用基因表达谱分析交互分析(GEPIA)确定乳腺癌和正常组织中的 PRR11 表达。通过 MTT 测定、克隆形成测定、划痕愈合测定、Transwell 测定和流式细胞术分别测量乳腺癌细胞的活力、增殖、迁移、侵袭和凋亡。通过定量实时聚合酶链反应(qRT-PCR)和 Western blot 检测 PRR11 和上皮-间充质转化(EMT)相关和凋亡相关(B 细胞淋巴瘤 2,Bcl-2;Bcl-2 相关蛋白 X,Bax)蛋白的表达。结果表明,随着超声强度的增加,MCF7 细胞的活力降低。GEPIA 的结果表明 PRR11 在乳腺癌中上调。US 介导的 PRR11 沉默比 Lipofectamine 3000 更有效。通过 Lipofectamine 3000 转染的 siPRR11 抑制细胞生长和转移,促进细胞凋亡。此外,E-钙粘蛋白(E-cad)和 Bax 表达较高,但 N-钙粘蛋白(N-cad)、Snail 和 Bcl-2 表达较低。然而,过表达的 PRR11 则产生相反的效果。更重要的是,与 Lipofectamine 3000 相比,使用 US 转染 siPRR11 和 PRR11 过表达质粒具有更高的疗效。US 转染 PRR11 siRNA 对抑制乳腺癌进展具有更好的效果。本研究结果为乳腺癌的治疗提供了新的思路。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cb04/7607193/ea1c4582b4bf/bsr-40-bsr20201854-g1.jpg

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