Differential gene expression profiles between SKOV3.ip1-S116A cells (Ser116 of PEA-15 substituted with alanine) and SKOV3.ip1-S116D cells (Ser116 of PEA-15 substituted with aspartic acid).

To dissect the molecular mechanisms of PEA-15-mediated paclitaxel sensitization in ovarian cancer cells, we performed cDNA microarray analysis using SKOV3.ip1-S116A cells (Ser116 of PEA-15 substituted with alanine) and SKOV3.ip1-S116D cells (Ser116 of PEA-15 substituted with aspartic acid). cDNA microarray data analysis showed that SCLIP (SCG10-like protein), also known as STMN3, was highly expressed in SKOV3.ip1-S116D cells and was involved in pPEA-15-mediated paclitaxel sensitization in ovarian cancer cells. SKOV3.ip1-S116A cells (Ser116 of PEA-15 substituted with alanine) and SKOV3.ip1-S116D cells (Ser116 of PEA-15 substituted with aspartic acid) were used. Total RNA was extracted and purified using RNeasy mini kit (Qiagen, Inc.) according to the manufacturer’s instructions. The integrity of the obtained RNA was assessed using an Agilent 2100 BioAnalyzer (Agilent Technologies). The Affymetrix HGU133 plus platform was used for hybridization, staining, and imaging of the arrays by following the manufacturer’s instructions. Gene expression analysis was performed in triplicate.

为解析PEA-15介导卵巢癌细胞对紫杉醇增敏的分子机制，本研究采用SKOV3.ip1-S116A细胞（PEA-15的Ser116位点突变为丙氨酸）与SKOV3.ip1-S116D细胞（PEA-15的Ser116位点突变为天冬氨酸）开展cDNA微阵列分析。cDNA微阵列数据分析结果显示，SCLIP（SCG10样蛋白，又名STMN3）在SKOV3.ip1-S116D细胞中呈高表达状态，且其参与了磷酸化PEA-15（pPEA-15）介导的卵巢癌细胞紫杉醇增敏过程。本研究所用细胞模型为SKOV3.ip1-S116A细胞（PEA-15的Ser116位点突变为丙氨酸）与SKOV3.ip1-S116D细胞（PEA-15的Ser116位点突变为天冬氨酸）。依照制造商说明书，采用RNeasy迷你试剂盒（Qiagen公司）提取并纯化总RNA；使用Agilent 2100生物分析仪（Agilent Technologies）评估所获总RNA的完整性。采用Affymetrix HGU133 plus芯片平台，按照厂商规程完成芯片的杂交、染色与成像。基因表达分析设置三次生物学重复。