Thériault Brigitte L, Cybulska Paulina, Shaw Patricia A, Gallie Brenda L, Bernardini Marcus Q
Campbell Family Cancer Research Institute, Ontario Cancer Institute, University Health Network, Toronto, ON, Canada.
Department of Obstetrics and Gynecology, University of Toronto, Toronto, ON, Canada.
J Ovarian Res. 2014 Dec 21;7:123. doi: 10.1186/s13048-014-0123-1.
Previously, it has been shown that KIF14 mRNA is overexpressed in ovarian cancer (OvCa), regardless of histological subtype. KIF14 levels are independently predictive of poor outcome and increased rates of recurrence in serous OvCa patients. Furthermore, it has been shown that KIF14 also controls the in vivo tumorigenicity of OvCa cell lines. In this study, we evaluate the potential of KIF14 as a therapeutic target through selective inhibition of KIF14 in primary high-grade serous patient-derived OvCa cells.
To assess the dependence of primary serous OvCa cultures on KIF14, protein levels in 11 prospective high grade serous ovarian cancer samples were increased (KIF14 overexpression by transfection) or decreased (anti-KIF14 shRNA) in vitro, and proliferative capacity, anchorage independence and xenograft growth were assessed.
Seven of eleven samples demonstrated increased/decreased in vitro proliferation in response to KIF14 overexpression/knockdown, respectively. When examining in vitro tumorigenicity (colony formation) and in vivo growth (subcutaneous xenografts) in response to KIF14 manipulation, none of the samples demonstrated growth in soft agar (11 samples), or xenograft growth (4 samples).
Although primary high-grade serous OvCa cells may depend on KIF14 for in vitro proliferation we were unable to demonstrate a role for KIF14 on tumorigenicity or develop an in vivo model for assessment. We have, however developed an effective in vitro method to evaluate the effect of target gene manipulation on the proliferative capacity of primary OvCa cultures.
此前研究表明，无论组织学亚型如何，KIF14 mRNA在卵巢癌（OvCa）中均过度表达。KIF14水平可独立预测浆液性OvCa患者的不良预后和复发率增加。此外，研究表明KIF14还控制OvCa细胞系的体内致瘤性。在本研究中，我们通过在原发性高级别浆液性患者来源的OvCa细胞中选择性抑制KIF14来评估KIF14作为治疗靶点的潜力。
为评估原发性浆液性OvCa培养物对KIF14的依赖性，在体外增加（通过转染使KIF14过表达）或降低（抗KIF14 shRNA）11份前瞻性高级别浆液性卵巢癌样本中的蛋白质水平，并评估增殖能力、锚定非依赖性和异种移植生长情况。
11份样本中有7份分别对KIF14过表达/敲低表现出体外增殖增加/减少。在检查对KIF14操作的体外致瘤性（集落形成）和体内生长（皮下异种移植）时，没有样本在软琼脂中生长（11份样本）或异种移植生长（4份样本）。
虽然原发性高级别浆液性OvCa细胞在体外增殖可能依赖KIF14，但我们无法证明KIF14在致瘤性方面的作用，也无法建立用于评估的体内模型。然而，我们开发了一种有效的体外方法来评估靶基因操作对原发性OvCa培养物增殖能力的影响。
