1Department of Gastroenterology,Affiliated Hospital of Zunyi Medical University; 2Department of Oncology; 3Department of General Medicine,Hospital of Zunyi Medical University, Zunyi 563000,China; 4Tumor Radiotherapy Department,95th Hospital of Liberation Army,Putian 351100,China
AbstractObjective To investigate the effect and mechanism of LncRNA ANRIL on the radiosensitivity of HCT116 cells line and nude mouse transplant tumors. Methods The expression of LncRNA ANRIL in colorectal cancer cells was detected by qPCR. The negative control siRNA,ANRIL siRNA,miR-NC mimic,miR-195 mimic,miR-NC inhibitor and miR-195 inhibitor were transfected into HCT116 cells,and marked as negative control group,silencing ANRIL group,overexpressing miR-NC group,overexpressing miR-195 group,inhibiting miR-NC group and inhibiting miR-195 group, and the HCT116 cells without any treatment were marked as the blank control group. The clone formation assay was used to detect radiosensitivity of colorectal cancer cells,flow cytometry was used to detect apoptosis. The web site,StarBase,was used to predict the downstream miRNAs of ANRIL and dual luciferase reporter gene assay was used to further verify. Subcutaneous tumor transplantation assay was used to detect the effect of ANRIL on the growth of colorectal cancer cells after irradiation. Results After irradiation with 2,4,6 and 8 Gy,the cell survival fraction of silencing ANRIL group was significantly decreased when compared with that of negative control group (P<0.05),and the radiosensitivity ratio was 1.52. The apoptosis rate of the silencing ANRIL+4 Gy group was significantly higher than that of the negative control+4 Gy group ((27.86±2.78)% vs. (12.06±1.46)%,P<0.05). The results of the experiment on nude mouse transplant tumors showed that the tumor volume in the negative control group was lower than that of the silent ANRIL group on days 13, 16, 19, 22 and 25 ((234±66) mm3,(273±63) mm3,(296±72) mm3,(321±85) mm3 and (403±94) mm3vs. (357±79) mm3,(485±124) mm3,(617±143) mm3,(764±174) mm3 and (985±221) mm3P<0.05). MiR-195 is a target gene of ANRIL,and inhibition of miR-195 can reverse the inhibitory effect of silencing ANRIL on radiosensitivity,apoptosis and xenografts of HCT116 cells. Conclusions LncRNA ANRIL regulates the radiosensitivity of colorectal cancer cells by miR-195,which may provide a new sensitizing target for clinical colorectal cancer radiotherapy.
Chen Xiaoyan,Wu Chenbin,Tian Xin et al. LncRNA ANRIL target miR-195 experimental study of radiation sensitivity of HCT116 cells and nude mouse transplant tumors[J]. Chinese Journal of Radiation Oncology, 2019, 28(11): 858-861.
Chen Xiaoyan,Wu Chenbin,Tian Xin et al. LncRNA ANRIL target miR-195 experimental study of radiation sensitivity of HCT116 cells and nude mouse transplant tumors[J]. Chinese Journal of Radiation Oncology, 2019, 28(11): 858-861.
[1] Luo J,Qu J,Wu DK,et al. Long non-coding RNAs:a rising biotarget in colorectal cancer[J]. Oncotarget,2017,8(13):22187-22202. DOI:10.18632/oncotarget.14728. [2] Fanale D,Castiglia M,Bazan V,et al. Involvement of non-coding RNAs in chemo-and radioresistance of colorectal cancer[J]. Adv Exp Med Biol,2016,937(2):207-228. DOI:10.1007/978-3-319-42059-2_11. [3] Hu X,Jiang H,Jiang X. Downregulation of LncRNA ANRIL inhibits proliferation,induces apoptosis,and enhances radiosensitivity in nasopharyngeal carcinoma cells through regulating miR-125a[J]. Cancer Biol Ther,2017,18(5):331-338. DOI:10.1080/15384047.2017.1310348. [4] Meseure D,Vacher S,Alsibai KD,et al. Expression of ANRIL-polycomb complexes-CDKN2A/B/ARF genes in breast tumors:identification of a Two-Gene (EZH2/CBX7) Signature with independent prognostic Value[J]. Mol Cancer Res,2016,14(7):623-633. DOI:10.1158/1541-7786. MCR-15-0418. [5] Yi L,Zhou X,Ling X,et al. Long noncoding RNA ANRIL could be transactivated by c-Myc and promote tumor progression of non-small-cell lung cancer[J]. Onco Targets Ther,2016,9:3077-3084. DOI:10.2147/OTT. S102658. [6] Sun Y,Zheng ZP,Li H,et al. ANRIL is associated with the survival rate of patients with colorectal cancer,and affects cell migration and invasion in vitro[J]. Mol Med Rep,2016,14(2):1714-1720. DOI:10.3892/mmr.2016.5409. [7] Zheng L,Chen J,Zhou Z,et al.miR-195 enhances the radiosensitivity of colorectal cancer cells by suppressing CARM1[J]. Oncol Targ Ther,2017,10:1027-1038. DOI:10.2147/OTT. S125067. [8] Quanquan S,Tongxin L,Yawei Y,et al. MiR-200c inhibits autophagy and enhances radiosensitivity in breast cancer cells by targeting UBQLN1[J]. Int J Cancer,2015,136(5):1003-1012. DOI:10.1002/ijc.29065. [9] 王启船,王青,屈中玉,等.miR-195在胃癌细胞中的表达及对癌细胞凋亡的机制研究[J]. 中国医药生物技术,2018,13(3):238-242. DOI:10.3969/j.issn.1673-713X.2018.03.008. Wang QC,Wang Q,Qu ZY,et al.miR-195 Expression in gastric cancer cells and the mechanism of apoptosis in cancer cells[J]. Chin Med Biotechnol,2018,13(3):238-242. DOI:10.3969/j.issn.1673-713X.2018.03.008. [10] Lin L,Lin C,Yingxin X,et al.microRNA-195 promotes apoptosis and suppresses tumorigenicity of human colorectal cancer cells[J]. Biochem Biophys Res Commun,2010,400(2):236-240. DOI:10.1016/j.bbrc.2010.08.046. [11] Militello G,Weirick T,John D,et al. Screening and validation of LncRNAs and circRNAs as miRNA sponges[J]. Br Bioinform,2017,18(5):780-788. DOI:10.1093/bib/bbw053.