Mechanisms of drug-resistance and apoptosis in oral carcinoma cells

口腔癌细胞耐药及凋亡机制

• 批准号: 16591893
• 负责人: OHBA Takeshi
• 金额: $ 2.18万
• 依托单位: Kyushu Dental College
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2004
• 资助国家: 日本
• 起止时间: 2004 至 2005
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-16591893/
• 关键词: drug-resistance cells; transcriptional factor; nuclear proteins; apoptosis; nucleorin; okadaic acid; cancer cells; protein dephosphorylation; 核内蛋白; ニュークレオフォスミン; AgNORs; 制癌剤; 口腔癌細胞; RNAi; 蛋白質脱リン酸化酵素

We examined the effects of anticancer drugs on induction of apoptosis and the transcriptional regulation of the multidrug resistance-1 (MDR1) gene in' oral squamous carcinoma cells. SCCKN cells were shown to be highly sensitive to various anticancer drugs, whereas SCCTF cells were minimally sensitive to these reagents. Peplomycin induced apoptosis in both SCCKN and SCCTF Cells in a dose-dependent fashion with the maximal effect at concentrations of 1 μM and 10 μM, respectively. Apoptosis was induced in SCCKN cells treated with 1 μM peplomycin, but not in SCCTF cells. RT-PCR revealed that MDR1 mRNA was highly expressed in SCCTF cells while it was under the limit of detection in SCCKN cells. After treatment with Hoechst 33342 to assess efflux action of MDR1 protein, intense staining occurred in SCCKN cells, whereas the staining was very weak in SCCTF cells. With an electrophoretic mobility shift assay using the MDR1 promoter region, a DNA-protein. Protein phosphorylation and dephosphorylation has been recognized as a key mechanism in cell function. Nucleolin is an abundantly expressed nucleolar phosphoprotein and is located mainly in the nucleolus. The staining pattern of nucleolin in cultured cells is similar to that of PP 1δ. Nucleolin was demonstrated to bind to PP1δ in nucleolus by using immunocytochemical and immunoprecipitation. methods. A major band, 110 kDa, was detected in the proteins obtained from the control cells. The level of the 110 kDa protein decreased in the apoptotic cells, whereas an additional band, 80 kDa, appeared and the level of this protein increased in the proteins prepared from okadaic acid-induced apoptotic cells. Our results indicate that PP1δ directly binds to nucleolin in the nucleolus and that nucleolin is cleaved during apoptosis.

我们检查了抗癌药对凋亡诱导的影响以及在口服鳞状癌细胞中多药抗性-1（MDR1）基因的转录调节。 SCCKN细胞被证明对各种抗癌药物高度敏感，而SCCTF细胞对这些试剂的敏感性最低。催产霉素以剂量依赖性的方式诱导SCCKN和SCCTF细胞的凋亡，分别在1μM和10μM下具有最大作用。在用1μM催眠霉素处理的SCCKN细胞中诱导凋亡，但在SCCTF细胞中诱导。 RT-PCR显示，MDR1 mRNA在SCCCTF细胞中高度表达，而在SCCKN细胞中的检测极限。用HOECHST 33342处理以评估MDR1蛋白的外排作用后，SCCKN细胞中发生了强烈的染色，而SCCTF细胞中的染色非常弱。使用MDR1启动子区域DNA-蛋白质的电泳移动性转移测定法。蛋白质磷酸化和去磷酸化已被认为是细胞功能的关键机制。核仁素是一种绝对表达的核磷蛋白，主要位于核olus中。核橄榄培养细胞的染色模式与PP1δ的染色模式相似。通过使用免疫细胞化学和免疫沉淀，核苷被证明与核仁中的PP1δ结合。方法。在从对照细胞获得的蛋白质中检测到一个主要带，110 kDa。凋亡细胞中110 kDa蛋白的水平降低，而另外一个80 kDa的带出现了，该蛋白质的水平在冈田酸诱导的凋亡细胞制备的蛋白中升高。我们的结果表明，PP1δ直接与核仁中的核苷结合，并且在凋亡过程中裂解了核仁素。

项目成果

Cleavage of nucleolin and AgNOR proteins during apoptosis induced by anti-cancer drugs in human salivary gland cells.

抗癌药物诱导人唾液腺细胞凋亡过程中核仁素和 AgNOR 蛋白的裂解。

• 发表时间: 2005
• 期刊: Journal of Oral Pathology & Medicine 34
• 作者: Ryoma Nakao;Hidenobu Senpuku;Haruo Watanabe;Tanaka T.;Inoue H;Morimoto Y;Ono K;Okabe S;Kito S;Kito S.;Morimoto Y.;Tanaka T.;Morimoto Y.;Tanaka T.;Kito S.
• 通讯作者: Kito S.

Okadaic acid induces tyrosine phosphorylation of IκBα that mediated by PKR pathway in human osteoblastic MG63 cells

• DOI: 10.1007/s11010-005-4440-y
• 发表时间: 2005-08-01
• 期刊: MOLECULAR AND CELLULAR BIOCHEMISTRY
• 影响因子: 4.3
• 作者: Morimoto, H;Ozaki, A;Haneji, T
• 通讯作者: Haneji, T

Association of matrix metelloproteinase (MMP)-1 promoter polymorphism with head and neck squamous cell carcinoma.

基质金属蛋白酶（MMP）-1启动子多态性与头颈鳞状细胞癌的关联。

• 发表时间: 2004
• 期刊: Cancer Letters 211
• 作者: Shunji.Izawa;Yoko.Tashiro;Junichi.Doi;Satoshi.Nakabo;Yoshihiro.Nishitani;Tomomasa.Takatsuka;Toshiyuki.Itota;Masahiro.Yoshiyama;Hashimoto T.;U.K.Bhawal;FujitaM.;Okamura H.;A.Otsubo;MorimotoH.;Morimoto H.;M.Sawajiri;Takeshi Ohba;Yasuaki Tate;U.K.Bhawal;Yasuhiro Morimoto;Nagako Yoshiba;Tomoko Hashimoto
• 通讯作者: Tomoko Hashimoto

Okadaic acid induces apoptosis through double-stranded RNA-dependent protein kinase/eukaryotic initiation factor-2α pathway in human osteoblastic MG63 cells

• DOI: 10.1093/jb/mvh144
• 发表时间: 2004-10-01
• 期刊: JOURNAL OF BIOCHEMISTRY
• 影响因子: 2.7
• 作者: Morimoto, H;Okamura, H;Haneji, T
• 通讯作者: Haneji, T

Evaluation of the relationship between periapical lesions/sclerotic bone and general bone density as a possible gauge of general health among 80 years old

评估根尖周病变/硬化骨与一般骨密度之间的关系，作为 80 岁老年人总体健康状况的可能衡量标准

• 发表时间: 2005
• 期刊: Oral Surg Oral Med Oral Pathol Oral Radiol Endod 99
• 作者: Nipaporn Wattanawongpitak;Takako Yoshikawa;Junji Tagami;Ohba T.
• 通讯作者: Ohba T.