切换至 "中华医学电子期刊资源库"

中华细胞与干细胞杂志(电子版) ›› 2023, Vol. 13 ›› Issue (03) : 151 -158. doi: 10.3877/cma.j.issn.2095-1221.2023.03.004

论著

干扰LINC00466通过miR-493-3p/MIF抑制子宫内膜癌RL95-2细胞恶性生物学行为
刘燕, 叶亚萍, 郑艳莉()   
  1. 226002 南通,江苏省南通市第二人民医院妇产科
  • 收稿日期:2023-03-22 出版日期:2023-06-01
  • 通信作者: 郑艳莉
  • 基金资助:
    南通市科技项目(MS12018007)

Interference with LINC00466 inhibits the malignant biological behavior of endometrial cancer RL95-2 cells through miR-493-3p/MIF

Yan Liu, Yaping Ye, Yanli Zheng()   

  1. Department of Obstetrics and Gynecology, the Second People's Hospital of Nantong, Nantong 226002, China
  • Received:2023-03-22 Published:2023-06-01
  • Corresponding author: Yanli Zheng
引用本文:

刘燕, 叶亚萍, 郑艳莉. 干扰LINC00466通过miR-493-3p/MIF抑制子宫内膜癌RL95-2细胞恶性生物学行为[J/OL]. 中华细胞与干细胞杂志(电子版), 2023, 13(03): 151-158.

Yan Liu, Yaping Ye, Yanli Zheng. Interference with LINC00466 inhibits the malignant biological behavior of endometrial cancer RL95-2 cells through miR-493-3p/MIF[J/OL]. Chinese Journal of Cell and Stem Cell(Electronic Edition), 2023, 13(03): 151-158.

目的

探讨LINC00466对子宫内膜癌RL95-2细胞生物学行为的影响及其作用机制。

方法

收集43例子宫内膜癌患者癌组织和癌旁组织,qRT-PCR法检测组织中LINC00466和miR-493-3p表达。体外培养RL95-2细胞,转染LINC00466小干扰RNA或miR-493-3p模拟物、或共转染LINC00466小干扰RNA与miR-493-3p抑制剂后,CCK-8法、Transwell实验分别检测细胞增殖抑制率、迁移和侵袭,qRT-PCR和Western blot分别检测细胞中巨噬细胞移动抑制因子(MIF) mRNA和蛋白表达。双荧光素酶报告基因实验对LINC00466和miR-493-3p调控关系进行验证。两组间比较采用t检验。

结果

子宫内膜癌组织中LINC00466的表达量较癌旁组织(2.66±0.34比0.95±0.21)升高,miR-493-3p的表达量较癌旁组织(0.42±0.10比0.98±0.15)降低,差异有统计学意义(P均< 0.001)。干扰LINC00466或过表达miR-493-3p增加RL95-2细胞增殖抑制率,减少细胞迁移数、侵袭数及细胞中MIF mRNA和蛋白表达(P < 0.001)。LINC00466靶向结合miR-493-3p,且干扰LINC00466的RL95-2细胞中miR-493-3p表达增加。敲减miR-493-3p逆转干扰LINC00466对RL95-2细胞增殖、迁移和侵袭及MIF表达的影响。

结论

LINC00466在子宫内膜癌组织中表达上调,干扰LINC00466可能通过靶向上调miR-493-3p和阻碍MIF表达来抑制子宫内膜癌RL95-2细胞增殖、迁移和侵袭。

Objective

To investigate the effect of LINC00466 on the biological behavior of endometrial cancer RL95-2 cells and its mechanism.

Methods

The cancer tissues and adjacent tissues of 43 patients with endometrial cancer were collected, and the expression of LINC00466 and miR-493-3p in the tissues were detected by qRT-PCR. RL95-2 cells were cultured in vitro and transfected with LINC00466 small interfering RNA or miR-493-3p mimic, or co-transfected with LINC00466 small interfering RNA and miR-493-3p inhibitor, and then CCK-8 method and Transwell assay were used to detect cell proliferation inhibition rate, migration and invasion. qRT-PCR and Western blot were used to detect the mRNA and protein expression of macrophage migration inhibitory factor (MIF) in cells, respectively. The regulatory relationship between LINC00466 and miR-493-3p was verified by the dual-luciferase reporter gene experiment. Comparison between the two groups was performed by t test.

Results

The expression of LINC00466 in endometrial cancer tissues was higher than that in adjacent tissues (2.66 ± 0.34 vs 0.95 ± 0.21) , but the expression of miR-493-3p was lower than that in adjacent tissues (0.42 ± 0.10 vs 0.98 ± 0.15) , and the differences were statistically significant (both P < 0.001) . Interfering with LINC00466 or overexpressing miR-493-3p increased the proliferation inhibition rate of RL95-2 cells, but decreased the number of cell migration and invasion and the mRNA and protein expression of MIF in cells (P < 0.001) . LINC00466 targets miR-493-3p, and the expression of miR-493-3p was increased in RL95-2 cells that interfered with LINC00466. Knockdown of miR-493-3p reversed the effects of interfering with LINC00466 on proliferation, migration, invasion and the expression of MIF in RL95-2 cells.

Conclusion

The expression of LINC00466 was up-regulated in endometrial cancer tissues. Interfering with LINC00466 may inhibit the proliferation, migration and invasion of endometrial cancer RL95-2 cells by targeting up-regulation of miR-493-3p and blocking MIF expression.

表1 引物序列信息
图1 子宫内膜癌组织中LINC00466和miR-493-3p表达注:aP < 0.001
图2 倒置显微镜下观察干扰LINC00466对RL95-2细胞迁移和侵袭的影响(结晶紫染色,×200)注:a、c图为si-NC组细胞的迁移和侵袭结果;b、d图为si-LINC00466组细胞的迁移和侵袭结果,可见被染色的细胞数与si-NC组相比减少
表2 干扰LINC00466对RL95-2细胞增殖、迁移和侵袭的影响( ± s
图3 LINC00466靶向结合miR-493-3p注:a图为预测的LINC00466和miR-493-3p的互补序列;b图为验证LINC00466和miR-493-3p靶向关系的双荧光素酶结果;aP < 0.001;ns为差异无统计学意义
图4 干扰LINC00466促进RL95-2细胞中miR-493-3p表达而抑制MIF表达注:a图为miR-493-3p的表达量;b图为MIF mRNA相对表达量;c图为干扰LINC00466对RL95-2细胞中MIF蛋白表达的影响;aP < 0.05
图5 倒置显微镜下观察过表达miR-493-3p对RL95-2细胞迁移、侵袭的影响(结晶紫染色,×200)注:a、c图为miR-NC组细胞的迁移和侵袭结果;b、d图为miR-493-3p组细胞的迁移和侵袭结果,可见被染色的细胞数与miR-NC组相比减少
表3 过表达miR-493-3p对RL95-2细胞增殖、迁移、侵袭的影响( ± s
图6 过表达miR-493-3p对RL95-2细胞中MIF表达的影响注:a图为MIF mRNA相对表达量;b图为MIF蛋白表达条带图和蛋白表达量;aP < 0.001
图7 倒置显微镜下观察敲减miR-493-3p逆转干扰LINC00466对RL95-2细胞迁移、侵袭的影响(结晶紫染色,×200)注:a、c图为si-LINC00466+anti-miR-NC组细胞的迁移和侵袭结果;b、d图为si-LINC00466+anti-miR-493-3p组细胞的迁移和侵袭结果,可见被染色的细胞数与si-LINC00466+anti-miR-NC组相比增多
图8 敲减miR-493-3p逆转干扰LINC00466对RL95-2细胞增殖、迁移、侵袭及MIF表达的影响注:a图为miR-493-3p相对表达量;b图为细胞抑制率;c图为细胞迁移数;d图为细胞侵袭数;e图为MIF mRNA相对表达量;f图为MIF蛋白表达条带图和蛋白表达量;aP < 0.001
1
Ma TG, Hu YB, Guo YX, et al. Tumor-promoting activity of long noncoding RNA LINC00466 in lung adenocarcinoma via miR-144-Regulated HOXA10 axis[J]. Am J Pathol, 2019, 189(11):2154-2170.
2
Li F, Shen ZZ, Xiao CM, et al. YY1-mediated up-regulation of lncRNA LINC00466 facilitates glioma progression via miR-508/CHEK1[J]. J Gene Med, 2021, 23(1):e3287-e3320.
3
Ding DX, Yang F, Chen ZJ, et al. Circ_0007385 regulates cell proliferation, apoptosis and stemness via targeting miR-493-3p/RAB22A axis in non-small cell lung cancer[J]. Thorac Cancer, 2022, 13(4):571-581.
4
Wang XY, Wang L, Xu PC, et al. LINC01605 promotes the proliferation of laryngeal squamous cell carcinoma through targeting miR-493-3p[J]. Eur Rev Med PharmacolSci, 2019, 23(23):10379-10386.
5
海啸,王琦,洪云飞. 血清肿瘤特异性生长因子、巨噬细胞移动抑制因子水平对骨肉瘤患者免疫靶向治疗效果的预测价值[J]. 癌症进展, 2022, 20(12):1267-1270.
6
高琳, 陈静, 戴云峰. 子宫内膜癌患者CerbB-2, NF-κB P50, MIF和IκBα蛋白表达水平与疾病严重程度的相关性研究[J]. 中国肿瘤临床与康复, 2020, 27(8):909-912.
7
Zhu W, Wu C, Zhou Z, et al. Acetate attenuates hyperoxaluria-induced kidney injury by inhibiting macrophage infiltration via the miR-493-3p/MIF axis[J]. Commun Biol, 2023, 6(1):270.
8
Zou C, Lv XZ, Wei HG, et al. Long non-coding RNA LINC00472 inhibits oral squamous cell carcinoma via miR-4311/GNG7 axis[J]. Bioengineered, 2022, 13(3):6371-6382.
9
Jian FF, Che XX, Zhang JJ, et al. The long-noncoding RNA SOCS2-AS1 suppresses endometrial cancer progression by regulating AURKA degradation[J]. Cell Death Dis, 2021, 12(4):351-363.
10
He YH, Xu SF, Qi Y, et al. Long noncoding RNA SNHG25 promotes the malignancy of endometrial cancer by sponging microRNA-497-5p and increasing FASN expression[J]. J Ovarian Res, 2021, 14(1):163-175.
11
Cong R, Kong FF, Ma J, et al. The PVT1/miR-612/CENP-H/CDK1 axis promotes malignant progression of advanced endometrial cancer[J]. Am J Cancer Res, 2021, 11(4):1480-1502.
12
武慧杰, 于桂芝, 李萌, 等. 基因间长链非编码RNA 113靶向微小RNA-493-3p对肺癌细胞侵袭和迁移的影响[J]. 临床肿瘤学杂志, 2021, 26(4):308-314.
13
Kleemann M, Schneider H, Unger K, et al. Induction of apoptosis in ovarian cancer cells by miR-493-3p directly targeting AKT2, STK38L, HMGA2, ETS1 and E2F5[J]. Cell Mol Life Sci, 2019, 76(3):539-559.
14
方瑜, 陈光伟, 杨洋, 等. 姜黄素在宫颈癌治疗中对MIF、VEGF-C、P53表达及肿瘤生长的影响[J]. 基因组学与应用生物学, 2018, 37(9):4148-4154.
15
成荣杰, 李楠. CD74介导巨噬细胞移动抑制因子对宫颈癌细胞生物学行为的影响[J]. 中国实验诊断学, 2020, 24(10):1689-1693.
16
Kong FY, Deng X, Kong XY, et al. ZFPM2-AS1, a novel lncRNA, attenuates the p53 pathway and promotes gastric carcinogenesis by stabilizing MIF[J]. Oncogene, 2018, 37(45):5982-5996.
17
Mamoori A, Gopalan V, Lu CT, et al. Expression pattern of miR-451 and its target (MIF) in colorectal cancer[J]. J ClinPathol, 2017, 70(4):308-312.
[1] 黄福, 王黔, 金相任, 唐云川. VEGFR2、miR-27a-5p在胃癌组织中的表达与临床病理参数及预后的关系研究[J/OL]. 中华普外科手术学杂志(电子版), 2024, 18(05): 558-561.
[2] 祝炜安, 林华慧, 吴建杰, 黄炯煅, 吴婷婷, 赖文杰. RDM1通过CDK4促进前列腺癌细胞进展的研究[J/OL]. 中华腔镜泌尿外科杂志(电子版), 2024, 18(06): 618-625.
[3] 赵蒙蒙, 黄洁, 余荣环, 王葆青. 过表达小GTP酶Rab32抑制非小细胞肺癌细胞侵袭性生长[J/OL]. 中华肺部疾病杂志(电子版), 2024, 17(04): 512-518.
[4] 王酉, 严斌, 狄文, 楼微华. 经脐单孔腹腔镜前哨淋巴结活检术在早期子宫内膜癌手术中的探讨[J/OL]. 中华腔镜外科杂志(电子版), 2024, 17(03): 173-176.
[5] 刘洪云, 李翠, 张海堂, 张玉英, 黄成香. 经脐单孔腹腔镜技术在子宫内膜癌手术中的应用[J/OL]. 中华腔镜外科杂志(电子版), 2024, 17(03): 182-184.
[6] 赵旭鹏, 王集琛, 田硕, 李宏召, 李修彬, 张旭. EP300 通过上调FKBP10 促进膀胱肿瘤细胞迁移和侵袭[J/OL]. 中华细胞与干细胞杂志(电子版), 2024, 14(05): 264-274.
[7] 黄程鑫, 陈莉, 刘伊楚, 王水良, 赖晓凤. OPA1 在乳腺癌组织的表达特征及在ER阳性乳腺癌细胞中的生物学功能研究[J/OL]. 中华细胞与干细胞杂志(电子版), 2024, 14(05): 275-284.
[8] 曾聿理, 雷发容, 肖慧, 邱德亮, 谢静, 吴寻. 氯普鲁卡因通过调控circRNA-ZKSCAN1表达抑制肝癌Huh-7细胞体外生长和转移的研究[J/OL]. 中华细胞与干细胞杂志(电子版), 2024, 14(04): 220-228.
[9] 李晶, 潘侠, 周芳, 汪晶, 洪佳. 普鲁卡因通过上调lncRNA DGCR5抑制胃癌细胞增殖、迁移和侵袭[J/OL]. 中华细胞与干细胞杂志(电子版), 2024, 14(03): 151-158.
[10] 刘杜先, 张杰东, 付鲁渝, 熊志强, 龚程, 张小雅, 高明悦, 孟俊宏, 刘兰侠. 沉默circXPO1抑制肝癌细胞恶性生物学行为[J/OL]. 中华细胞与干细胞杂志(电子版), 2024, 14(03): 159-166.
[11] 李博, 马秀岩, 孙杰. lncRNA TINCR对滋养层HTR-8/SVneo细胞生物学行为的影响及其机制[J/OL]. 中华细胞与干细胞杂志(电子版), 2024, 14(03): 167-172.
[12] 崔精, 鲍一帆, 沈晓明, 杨增辉, 高森, 鲍传庆. 结直肠癌中circMFSD12对肿瘤细胞功能及5-FU敏感性的调控[J/OL]. 中华结直肠疾病电子杂志, 2024, 13(04): 294-302.
[13] 王国强, 张纲, 唐建坡, 张玉国, 杨永江. LINC00839 调节miR-17-5p/WEE1 轴对结直肠癌细胞增殖、凋亡和迁移的影响[J/OL]. 中华消化病与影像杂志(电子版), 2024, 14(06): 491-499.
[14] 朱镭, 朱庆义. 金氏菌属:引起婴幼儿侵袭性传染病的新发病原体[J/OL]. 中华临床实验室管理电子杂志, 2024, 12(04): 229-237.
[15] 张芳芳, 李军, 赵玉洁, 于彤, 宁春平. 侵袭性血管黏液瘤的影像学特征并文献复习[J/OL]. 中华诊断学电子杂志, 2024, 12(04): 254-259.
阅读次数
全文


摘要