Department of Anesthesiology, Shandong Provincial Third Hospital, No.11, Wuyingshan Middle Road, Tianqiao District, Jinan, 250031, Shandong, China.
Department of Thoracic Surgery, Shandong ENT Hospital, Jinan, 250023, Shandong, China.
BMC Anesthesiol. 2021 Aug 30;21(1):210. doi: 10.1186/s12871-021-01429-z.
Opioid receptors are implicated in cell proliferation and cancer migration. However, the effects and underlying mechanisms of opioid receptor κ (OPRK1) in breast cancer remain unknown.
Small interfering RNA (siRNAs) was used to knockdown the expression of OPRK1. Western blot was used to determine the protein expression and reverse transcription-quantitative PCR (RT-qPCR) determined the genes transcription. Cell viability was detected by MTT assay and cell death rates were determined by Annexin V/PI and flow cytometry. Cell migration and invasion were detected by wound healing analysis and transwell assay, respectively.
Our research demonstrated that OPRK1 was overexpressed in breast cancer cells compared with the normal human mammary epithelial cells. OPRK1 knockdown could inhibited cell viability and migration in cancer cells, accompanied with the decreased proteins and genes expression of N-cadherin, Snail, MMP2 and Vimentin, while the E-cadherin expression was increased. Additionally, OPRK1 knockdown also promoted PI3K/AKT signaling inactivation. Activation of AKT reversed the OPRK1 knockdown-induced cell viability inhibition and migration suppression, while inhibition of AKT reduced cell viability and promoted cell death.
Our findings illustrated the role of OPRK1 played on promoting migration in vitro, and we also provided the therapeutic research of OPRK1 knockdown combined with AKT inhibition.
阿片受体参与细胞增殖和癌症迁移。然而，阿片受体 κ（OPRK1）在乳腺癌中的作用和潜在机制尚不清楚。
使用小干扰 RNA（siRNA）敲低 OPRK1 的表达。使用 Western blot 测定蛋白表达，逆转录定量 PCR（RT-qPCR）测定基因转录。通过 MTT 测定法检测细胞活力，通过 Annexin V/PI 和流式细胞术测定细胞死亡率。通过划痕愈合分析和 Transwell 测定分别检测细胞迁移和侵袭。
我们的研究表明，与正常的人乳腺上皮细胞相比，阿片受体 κ（OPRK1）在乳腺癌细胞中过表达。OPRK1 敲低可抑制癌细胞的活力和迁移，伴随着 N-钙粘蛋白、Snail、MMP2 和波形蛋白的蛋白和基因表达减少，而 E-钙粘蛋白的表达增加。此外，OPRK1 敲低还促进了 PI3K/AKT 信号通路失活。AKT 的激活逆转了 OPRK1 敲低诱导的细胞活力抑制和迁移抑制，而 AKT 的抑制减少了细胞活力并促进了细胞死亡。
我们的研究结果说明了 OPRK1 在促进体外迁移中的作用，并且我们还提供了 OPRK1 敲低与 AKT 抑制联合治疗的研究。
