title: [Long non-coding RNA influences radiosensitivity of colorectal carcinoma cell lines by regulating cyclin D1 expression].
abstract: OBJECTIVE
  To screen long non-coding RNA which influences radiosensitivity of colorectal carcinoma cell lines and investigate the mechanism.
  
  
  METHODS
  Under different doses of radiation, colony formation assay and single-hit multi-target model were conducted to draw dose-survival curve and SF2 value of colorectal carcinoma cell lines(RKO, Lovo) was calculated. High-throughput lncRNA/mRNA chips were used to screen lncRNA genes and protein coding genes with expression differences more than 2 folds between RKO, Lovo cell lines and RKO cell line receiving 2Gy radiation. The main action pathway was computed by Gene Ontology analysis combined with Pathway analysis in order to explore the mechanism which induces the effect of lncRNA on radiosensitivity of colorectal carcinoma cell lines. Further experiment on P53, P21, cyclin D1 expression contents of RKO cell line was confirmed by real-time RT-PCR.
  
  
  RESULTS
  Lovo(SF2=0.47) was more sensitivity to radiation than RKO(SF2=0.53) according to the outcome of colony formation assay. High-throughput lncRNA/mRNA chips identified a total of 268 lncRNA genes and 270 protein coding genes. Gene Ontology analysis showed that the expression of genes associated with cell cycle process were significantly different (38.6%). There was a significant relationship between expression of several lncRNAs and CCND1 gene. Real-time RT-PCR showed no significant differences of P53 and P21 expression in RKO and Lovo cell lines(P>0.05), while cyclin D1 expression of RKO cell line was higher than that of Lovo cell lines(P<0.05). After exposed to 2 Gy doses of radiation, there was an obvious decrease of cyclin D1 expression in RKO cell lines(P<0.05), while P53 and P21 expressions were not different(P>0.05).
  
  
  CONCLUSIONS
  The possible mechanism is that lncRNAs compose transcription compound to combine with CCND1 gene and influence radiosensitivity of colorectal carcinoma cell lines by regulating expression of cyclin D1, which is independent of P53-P21-cyclin D1 pathway.
authors.affiliation: Department of Colorectal Surgery, Changhai Hospital, The Second Military Medical University, Shanghai 200433, China.
authors.forename: Zhi-qi
authors.initials: ZQ
authors.lastname: Yu
authors.forename: Chang
authors.initials: C
authors.lastname: Zhang
authors.forename: Xin-yuan
authors.initials: XY
authors.lastname: Lao
authors.forename: Hao
authors.initials: H
authors.lastname: Wang
authors.forename: Xian-hua
authors.initials: XH
authors.lastname: Gao
authors.forename: Guang-wen
authors.initials: GW
authors.lastname: Cao
authors.forename: Wei-ping
authors.initials: WP
authors.lastname: Zhou
authors.forename: Chuan-gang
authors.initials: CG
authors.lastname: Fu
mesh_terms: D045744:Cell Line, Tumor; D015179:Colorectal Neoplasms / Q000378:metabolism* / Q000473:pathology; D019938:Cyclin D1 / Q000235:genetics* / Q000378:metabolism; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D062085:RNA, Long Noncoding; D011836:Radiation Tolerance
chemical_list: C530104:CCND1 protein, human; D062085:RNA, Long Noncoding; D019938:Cyclin D1