Third Military Medical University, Chongqing 400038, China; [corrected] Department of General Surgery, General Surgery Center of the PLA, General Hospital of Beijing Military Command, Beijing 100700, China.
Hepatobiliary Pancreat Dis Int. 2012 Aug 15;11(4):393-400. doi: 10.1016/s1499-3872(12)60197-0.
Phospholipid scramblase 1 (PLSCR1) not only participates in the transbilayer movement of phospholipids, but also plays a role in the pathogenesis and progression of cancers. The present study aimed to evaluate the effect of silencing PLSCR1 expression by RNA interference in colorectal cancer (CRC) and metastatic liver cancer.
The expression of PLSCR1 in CRC and metastatic liver cancer samples was assessed by immunohistochemistry. The cultured cells with the highest expression were selected for subsequent experiments. We designed three siRNA oligonucleotide segments targeted at PLSCR1. Successful transfection was confirmed. The biological behavior of the cells in proliferation, adhesion, migration and invasion was determined.
PLSCR1 protein expression increased significantly in the majority of CRC and metastatic liver cancer samples compared with normal samples. Lovo cells had the highest expression of PLSCR1. The siRNA-390 oligonucleotide segment had the best silencing effect. After transfection, Lovo cell proliferation was significantly inhibited compared with the controls in the MTT assay. Laminin and fibronectin adhesion assays showed Lovo cell adhesion was also significantly inhibited. In the migration assay, the number of migrating cells in the PLSCR1 siRNA-390 group was 50+/-12, significantly lower than the number in the siRNA-N group (115+/-28) and in the control group (118+/-31). In an invasion test, the number of invading cells in the PLSCR1 siRNA-390 group was 60+/-18, significantly lower than that in the siRNA-N group (97+/-26) and the control group (103+/-24).
PLSCR1 is overexpressed in CRC and metastatic liver cancer. Silencing of PLSCR1 by siRNA inhibits the proliferation, adhesion, migration and invasion of Lovo cells, which suggests that PLSCR1 contributes to the tumorigenesis and tumor progression of CRC. PLSCR1 may be a potential gene therapy target for CRC and associated metastatic liver cancer.
磷脂翻转酶 1（PLSCR1）不仅参与磷脂的跨膜运动，而且在癌症的发病机制和进展中发挥作用。本研究旨在评估 RNA 干扰沉默结直肠癌（CRC）和转移性肝癌中 PLSCR1 表达的效果。
通过免疫组织化学评估 CRC 和转移性肝癌样本中 PLSCR1 的表达。选择表达最高的培养细胞进行后续实验。我们设计了三个靶向 PLSCR1 的 siRNA 寡核苷酸片段。成功转染后，确定细胞的增殖、黏附、迁移和侵袭等生物学行为。
与正常样本相比，大多数 CRC 和转移性肝癌样本中 PLSCR1 蛋白表达显著增加。Lovo 细胞 PLSCR1 表达最高。siRNA-390 寡核苷酸片段具有最佳的沉默效果。与对照组相比，MTT 检测显示转染后 Lovo 细胞增殖明显受到抑制。层粘连蛋白和纤维连接蛋白黏附试验显示 Lovo 细胞黏附也明显受到抑制。在迁移试验中，PLSCR1 siRNA-390 组迁移的细胞数为 50+/-12，明显低于 siRNA-N 组（115+/-28）和对照组（118+/-31）。在侵袭试验中，PLSCR1 siRNA-390 组侵袭的细胞数为 60+/-18，明显低于 siRNA-N 组（97+/-26）和对照组（103+/-24）。
CRC 和转移性肝癌中 PLSCR1 表达上调。siRNA 沉默 PLSCR1 抑制 Lovo 细胞的增殖、黏附、迁移和侵袭，提示 PLSCR1 促进 CRC 的发生和发展。PLSCR1 可能成为 CRC 及其相关转移性肝癌的潜在基因治疗靶点。
