LncRNA PTENP1调控PTEN介导的乳腺癌转移

LncRNA PTENP1 modulates breast cancer metastasis by regulating PTEN expression

目的研究PTENP1与PTEN介导乳腺癌转移的分子机制,探索早期预测乳腺癌转移的分子靶标及临床乳腺癌治疗的新靶点。方法采用Real-time PCR技术方法检测乳腺癌组织及细胞株(MCF-7,MDA-MB-435)中PTENP1和PTEN表达情况;通过Real-time PCR、Western blot以及免疫荧光方法检测特异性调控PTENP1后PTEN表达量;采用CCK8实验及平板克隆实验,检测乳腺癌细胞株增殖能力;采用细胞划痕实验、Transwell侵袭实验,检测乳腺癌细胞株侵袭能力;通过绘制生存曲线,检测PTENP1及PTEN与临床乳腺癌患者预后的相关性。结果与乳腺癌癌旁组织及MCF-7细胞相比,PTENP1及PTEN在乳腺癌组织及MDA-MB-435细胞中低表达。特异性下调MCF-7细胞中PTENP1表达后,显著抑制PTEN的表达,而特异性上调MDA-MB-435中PTENP1表达后,PTEN的水平明显增高;特异性下调MCF-7细胞中PTENP1表达后,细胞的增殖、侵袭及转移能力显著增强,而特异性上调MDA-MB-435中PTENP1表达后,细胞的恶性程度部分抑制;通过生存曲线分析,高表达PTENP1及PTEN的乳腺癌患者5年生存时间显著高于低表达PTENP1及PTEN的乳腺癌患者。结论差异表达的PTENP1及PTEN与乳腺癌增殖、侵袭及转移特性密切相关,可能作为预测临床乳腺癌患者预后的潜在标志物。

Objective To investigate the possible mechanism of PTENP1 and PTEN mediated breast cancer metastasis,and explore a newly target point for early diagnosis and therapy in breast cancer.Methods The levels of PTENP1 and PTEN were detected in breast cancer tissues and cell lines(MCF-7,MDA-MB-435)by qRT-PCR.PTEN expression was determined by qRT-PCR,Western blot and immunofluorescence after modulation of PTENP1.The proliferative ability of breast cancer cells was measured by CCK8 assay and colony formation assay.Using wound healing assay and transwell assay,the aggressiveness of breast cancer cells was detected.Kaplan-Meier analysis was used to analyze the correlation between PTENP1/PTEN expression and the overall survival rate.Results The expression levels of PTENP1 and PTEN were lower in breast tumor and MDA-MB-435 cells than in adjacent non-tumor tissues and MCF-7 cells.Downregulation of PTENP1 suppressed PTEN level in MCF-7 cells,while overexpressed PTENP1 enhanced PTEN expression in MDA-MB-435 cells.The proliferation,migration and invasion abilities were significantly enhanced after decreasing PTENP1 level in MCF-7 cells.Overexpression of PTENP1 attenuated the malignancy of MDA-MB-435 cells.Breast cancer patients with higher PTENP1/PTEN levels lived longer than the patients with lower PTENP1/PTEN expression.Conclusion Differential levels of PTENP1 and PTEN were closely related to the proliferation,invasion and metastasis of breast cancer,both were potential biomarkers for predicting the prognosis of clinical breast cancer patients.