lncRNA ANCR对直肠癌细胞增殖和凋亡能力的影响及机制

doi:10.16689/j.cnki.cn11-9349/r.2020.03.017

摘要
图/表
参考文献
相关文章 (15)

全文: PDF (4059 KB) HTML (1 KB)
输出: BibTeX | EndNote (RIS)

摘要目的探讨长链非编码RNA（lncRNA）抗分化非编码RNA（ANCR）对结直肠癌（CRC）细胞生物学行为的影响及其作用机制。方法收集50例CRC患者的癌组织和癌旁组织，通过RT-qPCR检测组织中ANCR和叉头框蛋白O1（FOXO1）的表达水平并进行Pearson相关性分析。利用RNA 拉下和RNA免疫沉淀实验（RIP）确认ANCR是否可与FOXO1相互作用。将经过慢病毒包装的ANCR和FOXO1慢病毒质粒转染至人CRC细胞系SW480细胞中，细胞分为6组：对照组（未转染），NC组（转染慢病毒质粒空载），ANCR组（转染 pHRi-ANCR），sh-ANCR组（转染pHRi-sh-ANCR），FOXO1组（pHRi-FOXO1），sh-ANCR + sh-FOXO1组（同时转染pHRi-sh-ANCR和pHRi-sh-FOXO1）。RT-qPCR检测过表达和抑制ANCR后FOXO1的表达水平，5-乙炔基-2′脱氧尿嘧啶核苷（EdU）标记技术和流式细胞术分别检测各组细胞增殖、细胞周期和细胞凋亡情况。Western blot检测B淋巴细胞瘤-2（BCL-2）、BCL2相关蛋白X（BAX）、细胞周期蛋白D1（cyclin D1）及人P27蛋白（P27）的表达情况。结果与癌旁组织相比，CRC组织中ANCR表达明显增加，FOXO1表达明显减少（P<0.01），二者表达水平负相关（P<0.01）。ANCR能够结合并抑制FOXO1表达。与NC组相比，sh-ANCR组和FOXO1组细胞增殖数减少，细胞阻滞于G0/G1期，细胞凋亡数增多，BAX和P27增多，BCL-2和cyclin D1减少（P<0.05）。与sh-ANCR组相比，sh-ANCR + sh-FOXO1组上述指标得到逆转（P<0.05）。结论 lncRNA ANCR在CRC中表达升高并通过调控FOXO1的表达调控CRC细胞的细胞增殖和凋亡。

	服务

	把本文推荐给朋友
	加入我的书架
	加入引用管理器
	E-mail Alert
	RSS
	作者相关文章
	岳琰
	唐忠胜
	陈汇

Abstract：Objective To investigate the effect of long chain noncodingRNA (lncRNA) on the biological behavior of colorectal cancer (CRC) cells and its mechanism. Methods Cancer and adjacent tissues of 50 CRC patients were collected.The expression levels of ANCR and forkhead box protein O1 (FOXO1) in tissues were detected by RT-qPCR and Pearson correlation analysis was performed. RNA pull-down and RNA immunoprecipitation (RIP) experiments were used to confirm whether ANCR could interact with FOXO1. ANCR and FOXO1 lentivirus were transfected into CRC cell line SW480 cells. The cells were divided into the control group (untransfected), NC group (transfected with lentiviral vector), ANCR group (transfected with ANCR over-expressed lentivirus), sh-ANCR group (transfected with ANCR knockdown lentivirus), FOXO1group (transfected with FOXO1 overexpressing lentivirus), sh-ANCR + sh-FOXO1group (transfected with ANCR and FOXO1 simultaneously inhibited lentivirus). RT-qPCR was used to detect the expression of FOXO1 after over-expression and inhibition of ANCR. 5-ethynyl-2′-deoxyuridine (EdU) labeling technology and flow cytometry were used to detect cell proliferation, cell cycle and apoptosis in each group. Western blot was used to detect the expression of B-cell lymphoma-2 (BCL-2), BCL2-Associated X Protein (BAX), cycle proteins cyclin D1 (cyclin D1) and Human P27 protein (P27 protein). Results Compared with adjacent tissues, the expression of ANCR in CRC tissues increased significantly, while the expression of FOXO1 decreased significantly (P<0.01). The expression levels of the two were negatively correlated (P<0.01). ANCR is able to bind and inhibit FOXO1 expression. Compared with the NC group, the number of cells in the sh-ANCR group and FOXO1group was reduced, the cells were arrested in the G0 / G1 phase, the number of apoptosis was increased, BAX and P27 were increased, and BCL-2 and cyclin D1 were reduced (P<0.05). Compared with the sh-ANCR group, the above indexes of the sh-ANCR + sh-FOXO1 group were reversed (P<0.05). Conclusion LncRNA ANCR expression is increased in CRC and regulates cell proliferation and apoptosis of CRC cells by regulating FOXO1 expression.

Key words： Long noncoding RNA ANCR｜Forkhead box protein O1｜Colorectal cancer｜Cell proliferation｜Cell cycle｜Apoptosis

基金资助:绵阳市卫生健康委医学科研课题（201922）

通讯作者: 岳琰，电子邮箱：rlv25yb@163.com

引用本文:

岳琰，唐忠胜，陈汇. lncRNA ANCR对直肠癌细胞增殖和凋亡能力的影响及机制[J]. 肿瘤代谢与营养电子杂志, 2020, 7(3): 340-346.
Yue Yan,Tang Zhongsheng, Chen Hui. Effect of lncRNA ANCR on proliferation and apoptosis of colorectal cancer cells and its mechanism. Electron J Metab Nutr Cancer, 2020, 7(3): 340-346.

1.郭志娟, 吉茹. E-cadherin表达丢失与Vimentin表达在结直肠癌预后判断中的临床意义［J］. 实用肿瘤杂志, 2018, 33(2):181-184.
2.武雪亮, 王立坤, 黄先涛,等. 结直肠癌流行病学特征回顾性研究［J］. 中国医药导报, 2019, 16(20):60-63.
3.ZHONG W, QIANG C, BIN L, et al. Escin-induced DNA damage promotes escin-induced apoptosis in human colorectal cancer cells via p62 regulation of the ATM/γH2AX pathway［J］. Acta Pharmacol Sin, 2018, 39(10):1645-1660.
4.LI B, WANG Z, XIE J M, et al. TIGAR knockdown enhanced the anticancer effect of aescin via regulating autophagy and apoptosis in colorectal cancer cells［J］. Acta Pharmacol Sin, 2019, 40(1):111-121.
5.平贯芳, 熊万成, 邓智建. 长链非编码RNA浆细胞瘤转化迁移基因1在结直肠癌组织和细胞中的表达及临床意义［J］. 新乡医学院学报, 2019, 10(10),949-953.
6.LI R, WANG Y, XU Y, et al. Silencing the long noncoding RNA, TINCR, a molecular sponge of miR335, inhibits the malignant phenotype of epithelial ovarian cancer via FGF2 suppression［J］. Int J Oncol, 2019, 55(5):1110-1124.
7.YANG Z Y, YANG F, ZHANG Y L, et al. LncRNA-ANCR down-regulation suppresses invasion and migration of colorectal cancer cells by regulating EZH2 expression［J］. Cancer Biomark, 2017, 18(1):95-104.
8.朱金庚, 刘永, 涂志刚. LncRNA-ANCR介导PI3K及AKT蛋白对人胃癌细胞增殖迁移的作用机制［J］. 西部医学, 2019(9):1334-1338.
9.YUAN S, WANG J, YANG F, et al. Long noncoding RNA DANCR increases stemness features of hepatocellular carcinoma by derepression of CTNNB1［J］. Hepatology, 2016, 63(2):499-511.
10.DING X, WANG Q, TONG L, et al. Long non-coding RNA FOXO1 inhibits lung cancer cell growth through down-regulating PI3K/AKT signaling pathway［J］. Iran J Basic Med Sci, 2019, 22(5):491-498.
11.LIU B, ZHAO H, ZHANG L, et al. Silencing of long-non-coding RNA ANCR suppresses the migration and invasion of osteosarcoma cells by activating the p38MAPK signalling pathway［J］. BMC Cancer, 2019, 19(1):1112.
12.LI Z, HOU P, FAN D, et al. The degradation of EZH2 mediated by lncRNA ANCR attenuated the invasion and metastasis of breast cancer［J］. Cell Death Differ, 2017, 24(1):59.
13.FU T, COULTER S, YOSHIHARA E, et al. FXR regulates intestinal cancer stem cell proliferation［J］. Cell, 2019, 176(5):1098-1112. e18.
14.张新明, 郭德迎, 何海填,等. 肿瘤相关巨噬细胞对膀胱癌细胞增殖和血管生成的影响［J］. 现代肿瘤医学, 2018, 26(24):31-34.
15.罗娟, 周晓, 程志祥. FOXO1在非小细胞CRC中的研究进展［J］. 南京医科大学学报 (自然科学版), 2018, 38(8):1161-1165.
16.YUN CHEOL C, JI YOUNG K, WOO P J, et al. FOXO1 degradation via G9a-mediated methylation promotes cell proliferation in colon cancer［J］. Nucleic Acids Res, 2019, 47(4):1692-1705.
17.YAN T, HU G, WANG A, et al. Paris saponin Ⅶ induces cell cycle arrest and apoptosis by regulating Akt/MAPK pathway and inhibition of P-glycoprotein in K562/ADR cells［J］. Phytother Res, 2018, 32(5):898-907.
18.ZHOU Z, CHEN G, DENG C, et al. TCF19 contributes to cell proliferation of non-small cell lung cancer through inhibiting FOXO1［J］. Cell Biol Int, 2019, 43(12):1416-1424.
19.GAO Z, LIU R, YE N, et al. FOXO1 inhibits tumor cell migration via regulating cell surface morphology in non-small cell lung Cancer cells［J］. Cell Physiol Biochem, 2018, 48(1):138-148.
20.ZHU X, WU Y B, ZHOU J, et al. Upregulation of lncRNA MEG3 promotes hepatic insulin resistance via increasing FOXO1 expression［J］. Biochem Bioph Res Co, 2015, 469(2):319-325.
21.TEMIZ P, AKKAS G, NESE N, et al. Determination of apoptosis and cell cycle modulators (p16, p21, p27, p53, BCL-2,Bax, BCL-xL, and cyclin D1) in thyroid follicular carcinoma, follicular adenoma,and adenomatous nodules via a tissue microarray method［J］.Turk J Med Sci, 2015, 45(4):865-871.
22.黄虎, 栗娜, 杨柳,等. 非小细胞肺癌组织中NF-κB和CyclinD1及p27表达与患者化疗敏感性［J］. 贵阳医学院学报, 2019, 44(2):195-199.
23.刘蕾, 李砚, 王莎莎. Bcl-2与肿瘤转移的研究进展［J］. 中国医药导报, 2018, 15(7):40-43.
24.ZHANG F, PENG H. LncRNA-ANCR regulates the cell growth of osteosarcoma by interacting with EZH2 and affecting the expression of p21 and p27［J］. J Orthop Surg Res, 2017, 12(1):103.
25.SUN B, LI J, SHAO D, et al. Adipose tissue-secreted miR-27a promotes liver cancer by targeting FOXO1 in obese individuals［J］. Oncotargets Ther, 2015, 8:735-744.