shRNA 介导的 Slc38a1 沉默抑制人胰腺癌细胞的迁移，但不抑制其侵袭。
shRNA-mediated Slc38a1 silencing inhibits migration, but not invasiveness of human pancreatic cancer cells.
机构信息
Department of Integrative Oncology, Fudan University Shanghai Cancer Center; Department of Oncology, Shanghai Medical College, Fudan University, Shanghai 200032, China;
出版信息
Chin J Cancer Res. 2013 Oct;25(5):514-9. doi: 10.3978/j.issn.1000-9604.2013.09.03.
OBJECTIVE
Early metastasis is a major biological feature of pancreatic cancer. The current study examined whether silencing Slc38a1, a gene involved in energy metabolism, using short hairpin RNA (shRNA) could inhibit the growth, migration, and invasiveness of pancreatic cancer cells.
METHODS
A series of Slc38a1 shRNAs were designed and cloned into the pGPU6/GFP/Neo vectors. An shRNA with the most efficacious inhibitory action on SCL38A1 expression (65% inhibition) upon screening in DH5α bacteria was used to transfect SW1990 human pancreatic cancer cells. Cell growth, migration, and invasiveness were examined using cell counting kit-8, Boyden chamber without and with Matrigel, respectively.
RESULTS
Transfection of SW1990 cells with the SLCs38A1 shRNA significantly decreased the proliferation (P<0.0001) and migratory potential (by 46.7%, P=0.0399) of the cancer cells. Invasiveness, however, was not affected.
CONCLUSIONS
Inhibiting Slc38a1 using shRNA technology could decrease the growth and migration of representative pancreatic cancer cells. However, the fact that invasiveness was not affected suggested that SLC38A1 is unlikely to be responsible for early metastasis.
目的
早期转移是胰腺癌的主要生物学特征。本研究通过短发夹 RNA(shRNA)沉默参与能量代谢的基因 Slc38a1，来检测其是否能抑制胰腺癌细胞的生长、迁移和侵袭。
方法
设计并克隆了一系列 Slc38a1 shRNA 到 pGPU6/GFP/Neo 载体中。在 DH5α 细菌中筛选出对 SCL38A1 表达抑制作用最有效的 shRNA（抑制率为 65%），用于转染 SW1990 人胰腺癌细胞。通过细胞计数试剂盒-8、Boyden 室（无 Matrigel 和有 Matrigel）分别检测细胞生长、迁移和侵袭。
结果
SW1990 细胞转染 SLCs38A1 shRNA 后，癌细胞的增殖（P<0.0001）和迁移能力（降低 46.7%，P=0.0399）显著下降。然而，侵袭力不受影响。
结论
利用 shRNA 技术抑制 Slc38a1 可以降低代表性胰腺癌细胞的生长和迁移。然而，侵袭力不受影响这一事实表明，SLC38A1 不太可能是早期转移的原因。
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