合作案例

合作案例

项目简介

Long noncoding RNA AFAP1-AS1 predicts a poor prognosis and regulates non-small cell lung cancer cell proliferation by epigenetically repressing p21 expression
长链非编码RNA AFAP1-AS1预测不良预后,并且通过表观抑制p21表达从而调节非小细胞肺癌细胞增殖

服务内容
本文主要研究的是LncRNA通过表观遗传调控下游靶基因的表达进而影响疾病的发生。涉及的实验方法有:qPCR、MTT、细胞克隆、细胞周期、裸鼠成瘤、免疫组化、RNA-seq、RIP以及Chip-qPCR等。本文的研究采取了一般的研究思路三部曲:首先确定通过预后筛选目的基因,然后确定目的在疾病中的功能,最后研究其起功能的作用机制。研究结果最终表明:AFAP1-AS1高表达预示着非小细胞肺癌患者不良预后,并且,AFAP1-AS1通过表观遗传学抑制P21的表达,进而影响非小细胞肺癌细胞增殖。
交付标准

1、课题设计方案 

2、立题报告 

3、完整实验报告 

4、原始数据 

5、结题资料

结果展示

Figure1:ENSG00000231881在结肠癌临床组织样本中的表达水平检测



(A) Expression of lncRNA ENSG00000231881 in CRC and normal tissues. (B) Expression of lncRNA ENSG00000231881 at different stages in CRC tissues. (C) Survival curve based on lncRNA ENSG00000231881 expression level in CRC patients. (D) Expression levels of lncRNA ENSG00000231881 in clinical CRC tissue and paracancerous tissue samples obtained from 20 patients with CRC, as evaluated with qPCR assay. *P < 0.05 and **P < 0.01.



Figure2.ENSG00000231881在HCT-116、Lovo中的功能学研究



(A) Successful establishment of models for the overexpression and interference expression of ENSG00000231881, as evaluated by qPCR assay. (B) Effect of ENSG00000231881 expression level on cell proliferation, as observed with the MTT assay. (C) Effect of ENSG00000231881 expression level on cell metastasis, as observed in the transwell migration assay. (D) Effect of ENSG00000231881 expression level on the angiogenesis of HLECs, as evident in the tube formation experiment. (E) Effect of ENSG00000231881 expression on VEGFC mRNA expression level, as evaluated by qPCR assay. (F) Effect of ENSG00000231881 expression on VEGFC protein expression level, as evident in the western blot assay. **P < 0.01.


Figure3.ENSG00000231881与miR-133b结合作用验证



(A) Expression level of miR-133b with ENSG00000231881 modelling, as observed in qPCR assay. (B) Illustration of the binding points of lncRNA and miR-133b, http://www.mircode.org/. (C) Binding relationship between ENSG00000231881 and miR-133b, as verified by the dual luciferase reporter assay. *P < 0.05, **P < 0.01.



Figure4.miR-133b过表达可抑制ENSG00000231881过表达所引起的各种功能学改变



(A) Effect of miR-133b mimics on the expression levels of ENSG00000231881 and miR-133b in the ENSG00000231881 overexpression model, as evaluated by qPCR assay. (B) Effect of miR-133b mimics on cell proliferation in the ENSG00000231881 overexpression model, as evaluated by the MTT assay. (C) Effect of miR-133b mimics on cell metastasis in the ENSG00000231881 overexpression model, as observed in the transwell migration assay. (D) Effect of miR-133b mimics on the angiogenesis of HLECs in the ENSG00000231881 overexpression model, as seen in the tube formation experiment. (E) Effect of miR-133b mimics on the expression levels of mRNA and protein of VEGFC in the ENSG00000231881 overexpression model, as evaluated with qPCR and western blot analyses, respectively. **P < 0.01

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