Cannon Claire M, Trembley Janeen H, Kren Betsy T, Unger Gretchen M, O'Sullivan M Gerard, Cornax Ingrid, Modiano Jaime F, Ahmed Khalil
Am J Vet Res. 2017 Aug;78(8):946-953. doi: 10.2460/ajvr.78.8.946.
OBJECTIVE To investigate protein kinase CK2 (CK2) expression in squamous cell carcinoma (SCC) of cats and to examine effects of CK2 downregulation on in vitro apoptosis and viability in SCC. SAMPLE Biopsy specimens of oral mucosa and testis and blood samples from clinically normal cats, biopsy specimens of oral SCC from cats, and feline SCC (SCCF1) and mammary gland carcinoma (K12) cell lines. PROCEDURES Immunohistochemical labeling for CK2α was performed on biopsy specimens. Sequences of the CK2α subunit gene and CK2α' subunit gene in feline blood and feline cancer cell lines were determined by use of PCR and reverse-transcription PCR assays followed by direct Sanger sequencing. Specific small interfering RNAs (siRNAs) were developed for feline CK2α and CK2α'. The SCCF1 cells were treated with siRNA and assessed 72 hours later for CK2α and CK2α' expression and markers of apoptosis (via western blot analysis) and for viability (via 3-[4,5-dimethylthiazol-2-yl]-5-[3-carboxymethoxyphenyl]-2-[4-sulfophenyl]-2H-tetrazolium assays). RESULTS CK2α was expressed in all feline oral mucosa samples and 7 of 8 oral SCC samples. Expression of CK2α and CK2α' was successfully downregulated in SCCF1 cells by use of siRNAs, which resulted in decreased viability and induction of apoptosis. CONCLUSIONS AND CLINICAL RELEVANCE In this study, CK2 appeared to be a promising therapeutic target for SCCs of cats. A possible treatment strategy for SCCs of cats would be RNA interference that targets CK2.
目的 研究蛋白激酶CK2(CK2)在猫鳞状细胞癌(SCC)中的表达,并检测CK2下调对SCC体外凋亡和活力的影响。 样本 来自临床正常猫的口腔黏膜和睾丸活检标本及血液样本、猫口腔SCC活检标本、猫SCC(SCCF1)和乳腺癌(K12)细胞系。 步骤 对活检标本进行CK2α免疫组织化学标记。采用聚合酶链反应(PCR)和逆转录PCR检测法,随后进行直接桑格测序,测定猫血液和猫癌细胞系中CK2α亚基基因和CK2α'亚基基因的序列。针对猫CK2α和CK2α'开发了特异性小干扰RNA(siRNA)。用siRNA处理SCCF1细胞,72小时后评估CK2α和CK2α'的表达、凋亡标志物(通过蛋白质印迹分析)和活力(通过3-[4,5-二甲基噻唑-2-基]-5-[3-羧甲氧基苯基]-2-[4-磺基苯基]-2H-四唑鎓检测)。 结果 CK2α在所有猫口腔黏膜样本和8个口腔SCC样本中的7个中表达。通过使用siRNA成功下调了SCCF1细胞中CK2α和CK2α'的表达,这导致活力降低并诱导凋亡。 结论及临床意义 在本研究中,CK2似乎是猫SCC的一个有前景的治疗靶点。针对猫SCC的一种可能治疗策略是靶向CK2的RNA干扰。