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. 2018 Jun;28(6):693-695.
doi: 10.1038/s41422-018-0033-7. Epub 2018 Apr 8.

Circular RNA F-circEA produced from EML4-ALK fusion gene as a novel liquid biopsy biomarker for non-small cell lung cancer

Shuangyan Tan  1 , Qiheng Gou  1 , Wenchen Pu  1 , Chenglin Guo  1 , Yun Yang  2 , Ke Wu  1 , Yaxin Liu  1 , Lunxu Liu  1 , Yu-Quan Wei  1 , Yong Peng  3
Affiliations

Circular RNA F-circEA produced from EML4-ALK fusion gene as a novel liquid biopsy biomarker for non-small cell lung cancer

Shuangyan Tan et al. Cell Res. 2018 Jun.
No abstract available

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Conflict of interest statement

The authors declare no competing interests.

Figures

Fig. 1
Fig. 1
Identification and functional analysis of F-circEA in NSCLC. a Schematic representation of F-circEA generated from EML4-ALK fusion gene. The convergent primers (F1/R1) were used to detect EML4-ALK fusion site, and the divergent primer sets (F2/R2, F3/R3) were used to detect F-circEA. Violet curve represents siRNA location. b Agarose gel electrophoresis and Sanger sequencing of RT-PCR products from H2228 cells with F1/R1 primers, the arrow indicates EML4-ALK fusion site. c Identification of F-circEA in H2228 cells by RT-PCR with F2/R2 primers and Sanger sequencing. The arrow indicates F-circEA junction site. d Detection of F-circEA and EML4-ALK mRNA by RNA solution hybridization assay using 32P-labeled oligonucleotide probes crossing junction site and the fusion site. e Cell nucleus/cytoplasm fractionation and RT-qPCR analysis revealed the cytoplasmic distribution of F-circEA in H2228 cells. GAPDH mRNA and U6 snRNA represent cytoplasmic and nuclear RNA, respectively. Western blotting confirmed the efficiency of nucleus/cytoplasm isolation. Data are shown as the mean ± SD. f Knockdown efficiency of F-circEA siRNAs in H2228 cells analyzed by RT-qPCR. g F-circEA knockdown inhibited cell migration and invasion in H2228 cells. h Schematic representation of F-circEA-expressing plasmid with the reverse repeat of cirR-7 exons plus up-stream and down-stream flanking introns to facilitate RNA circularization. i RNA solution hybridization assays for F-circEA and EML4-ALK mRNA in H2228, H1299, empty vector-transfected (Ctrl-H1299), and F-circEA-overexpressing (F-circEA-H1299) H1299 cells. j Cell nucleus/cytoplasm fractionation and RT-qPCR analysis demonstrated that F-circEA was predominantly located in the cytoplasm of F-circEA-overexpressing H1299 cells. Data are shown as the mean ± SD. k Ectopic expression of F-circEA increased the migratory and invasive ability of H1299 cells. Results of statistical analysis are shown in the right panel. l RT-qPCR analysis showed effective knockdown of F-circEA by siRNA in F-circEA-overexpressing H1299 cells. The scramble siRNA (SCR) was used as negative control. m, n F-circEA knockdown attenuated the enhanced migratory and invasive ability of F-circEA-overexpressing H1299 cells. *P < 0.05. o, p Agarose gel electrophoresis and Sanger sequencing of RT-PCR products from tumor tissues (o) or plasma (p) of NSCLC patients with or without EML4-ALK fusion gene
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