Study on the mechanism of MALAT1 targeting miR-142-3p in ovarian cancer chemotherapy resistance

Abstract

Abstract:

ObjectiveTo study the mechanism of metastasis associated lung adenocarcinoma transcript 1 (MALAT1) targeting microRNA-142-3p (miR-142-3p) in ovarian cancer chemotherapy resistance.MethodsA total of 80 ovarian cancer tissues and paired normal tissues were collected in Shaanxi Provincial People's Hospital from February 2016 to February 2019. The relative expression levels of MALAT1 and miR-142-3p in ovarian cancer tissues and paired normal tissues were detected by real-time fluorescence quantitative polymerase chain reaction (PCR), and the correlation between MALAT1 and miR-142-3p was analyzed. The effects of abnormal expressions of MALAT1 and miR-142-3p on proliferation and chemotherapy sensitivity of 5-fluorouracil (5-FU) and cisplatin of ovarian cancer Hey cells were verified by CCK-8 assay. Dual luciferase reporter gene experiment was used to detect the targeted relationship between miR-142-3p and MALAT1 (Hey cells were divided into four groups: MALAT1 wt, MALAT1 wt+miR-142-3p mimic, MALAT1 mut, MALAT1 mut+miR-142-3p mimic). RNA immunoprecipitation assay was use to confirm the binding site of MALAT1 and miR-142-3p.ResultsIn the ovarian cancer tissues and paired normal tissues, the relative expression levels of MALAT1 were 0.000 52 (0.002 56) and 0.000 47 (0.000 89), with a statistically significant difference (Z=2.365, P=0.018); the relative expression levels of miR-142-3p were 0.001 19 (0.002 69) and 0.001 61 (0.008 48), with a statistically significant difference (Z=2.935,P=0.003). The relative expression level of MALAT1 was negatively correlated with miR-142-3p in the ovarian cancer tissues (r=-0.474,P<0.001). The relative expression level of miR-142-3p in the miR-142-3p mock group was statistically lower than that of MALAT1+miR-142-3p mimic group (0.004 18±0.001 24vs.0.006 51±0.000 28;t=3.174,P=0.017). The relative fluorescence concentrations of MALAT1 wt group and MALAT1 wt+miR-142-3p mimic group were 2.27±0.86 and 31.10±6.05 respectively, with a statistically significant difference (t=8.172,P<0.001). After 48, 72 and 96 hours of ovarian cancer Hey cells being transfected with MALAT1 overexpression plasmid, the absorbance (A) values of cells in the MALAT1 overexpression group were significantly greater than those in the control group (0.522±0.021vs.0.433±0.021; 0.644±0.012vs.0.544±0.051; 0.887±0.055vs.0.698±0.042), with statistically significant differences (allP<0.05). After MALAT1 being overexpressed in Hey cells, at 0.10 ng/μl concentration of 5-FU, the proliferation rate of cells in the overexpression group was significantly faster than that in the control group (0.615±0.036vs.0.506±0.042;t=4.432,P=0.002), and the cells at 1.00, 10.00, 100.00 ng/μl concentrations of 5-FU showed the same trends (allP<0.05). At 0.01 ng/μl concentration of cisplatin, the proliferation rate of cells in the overexpression group was significantly faster than that in the control group (0.777±0.015vs.0.733±0.039;t=2.355,P=0.023), and the cells at 0.10, 1.00, 10.00, 100.00 ng/μl concentrations of cisplatin showed the same trends (allP<0.05). After miR-142-3p being overexpressed in Hey cells, at 0.10 ng/μl concentration of 5-FU, the proliferation rate of cells in the overexpression group was significantly slower than that in the control group (0.512±0.051vs.0.744±0.119;t=4.028,P=0.004), and the cells at 1.00, 10.00, 100.00 ng/μl concentrations of 5-FU showed the same trends (allP<0.05). At 0.10 ng/μl concentration of cisplatin, the proliferation rate of cells in the overexpression group was significantly slower than that in the control group (0.520±0.043vs.0.674±0.096;t=3.441,P=0.009), and the cells at 1.00, 10.00, 100.00 ng/μl concentrations of cisplatin showed the same trends (allP<0.05). After ovarian cancer Hey cells being treated with 0.10, 1.00, 10.00, 100.00 ng/μl concentrations of 5-FU and cisplatin, the proliferation rates of cells in the MALAT1 overexpression group, MALAT1+miR-142-3p group and control group showed statistically significant differences (allP<0.05). Further pairwise comparisons revealed that the proliferation rates of cells in the MALAT1+miR-142-3p group were significantly slower than those in the MALAT1 overexpression group (allP<0.05).ConclusionMALAT1 can reduce the sensitivity of ovarian cancer cells to 5-FU and cisplatin by targeted miR-142-3p, leading to chemotherapy resistance of ovarian cancer.

Key words:Ovarian neoplasms,Fluorouracil,Cisplatin,Metastasis associated lung adenocarcinoma transcript 1,MicroRNA-142-3p

Fan Yangyang, Wang Ying, Yuan Feng, Du Shanping, He Rongrong, Jia Yan. Study on the mechanism of MALAT1 targeting miR-142-3p in ovarian cancer chemotherapy resistance[J]. Journal of International Oncology, 2020, 47(2): 82-89.

Figures/Tables10

组别	5-氟尿嘧啶浓度(ng/μl)
MALAT1过表达组	0.893±0.018	0.615±0.036	0.451±0.041	0.376±0.089	0.282±0.067
对照组	0.881±0.039	0.506±0.042	0.228±0.017	0.124±0.051	0.076±0.031
t值	0.625	4.432	11.143	5.481	6.242
P值	0.275	0.002	<0.001	<0.001	<0.001

组别	顺铂浓度(ng/μl)
MALAT1过表达组	0.777±0.015	0.510±0.030	0.337±0.031	0.280±0.066	0.197±0.013
对照组	0.733±0.039	0.456±0.042	0.228±0.017	0.124±0.051	0.067±0.027
t值	2.355	2.372	6.865	4.418	9.626
P值	0.023	0.045	<0.001	0.003	<0.001

组别	5-氟尿嘧啶浓度(ng/μl)
miR-142-3p过表达组	0.683±0.107	0.512±0.051	0.306±0.058	0.162±0.043	0.099±0.052
对照组	0.750±0.075	0.744±0.119	0.552±0.068	0.455±0.046	0.228±0.086
t值	1.147	4.028	6.198	10.348	2.868
P值	0.858	0.004	<0.001	<0.001	0.021

组别	顺铂浓度(ng/μl)
miR-142-3p过表达组	0.701±0.071	0.520±0.043	0.219±0.092	0.169±0.0038	0.128±0.029
对照组	0.821±0.133	0.674±0.096	0.556±0.107	0.437±0.065	0.271±0.041
t值	1.780	3.441	5.363	8.007	6.277
P值	0.935	0.009	<0.001	0.003	<0.001

组别	5-氟尿嘧啶浓度(ng/μl)
MALAT1过表达组	0.778±0.055	0.723±0.035^ab	0.596±0.063^ab	0.494±0.046^ab	0.418±0.053^ab
MALAT1+miR-142-3p组	0.735±0.098	0.635±0.040^b	0.458±0.062^b	0.351±0.064^b	0.233±0.057^b
对照组	0.620±0.073	0.413±0.064	0.280±0.064	0.188±0.043	0.114±0.035
F值	1.528	55.700	31.952	43.890	48.666
P值	0.322	<0.001	<0.001	<0.001	<0.001

组别	顺铂浓度(ng/μl)
MALAT1过表达组	0.794±0.073	0.753±0.044^ab	0.581±0.080^ab	0.481±0.007^ab	0.362±0.061^ab
MALAT1+miR-142-3p组	0.719±0.108	0.635±0.040^b	0.458±0.062^b	0.315±0.045^b	0.263±0.094^b
对照组	0.698±0.053	0.439±0.033	0.294±0.070	0.230±0.036	0.106±0.067
F值	1.235	81.417	20.618	72.654	14.787
P值	0.201	<0.001	<0.001	<0.001	<0.001

References21

[1]	Chen W, Sun K, Zheng R , et al. Cancer incidence and mortality in China, 2014[J]. Chin J Cancer Res, 2018,30(1):1-12. DOI: 10.21147/j.issn.1000-9604.2018.01.01.
[2]	Siegel RL, Miller KD, Jemal A . Cancer statistics, 2019[J]. CA Cancer J Clin, 2019,69(1):7-34. DOI: 10.3322/caac.21551.
[3]	郑荣寿, 孙可欣, 张思维 , 等. 2015年中国恶性肿瘤流行情况分析[J]. 中华肿瘤杂志, 2019,41(1):19-28. DOI: 10.3760/cma.j.issn.0253-3766.2019.01.005.
[4]	黄志有, 马必东, 黄兆明 . 复发性卵巢上皮癌女性患者无瘤生存状况及其影响因素分析[J]. 中国妇幼健康研究, 2017,28(7):788-789, 896. DOI: 10.3969/j.issn.1673-5293.2017.07.012.
[5]	王冉冉, 董珊珊, 龙瀛 , 等. HMMR-AS1介导EMT在上皮性卵巢癌顺铂获得性耐药中的作用[J]. 肿瘤药学, 2018,8(4):498-504. DOI: 10.3969/j.issn.2095-1264.2018.04.04.
[6]	李亚芳, 谷春会 . 复发性卵巢癌治疗进展[J]. 吉林医学, 2018,39(1):169-172.
[7]	Wu L, Zhang L, Zheng S . Role of the long non-coding RNA HOTAIR in hepatocellular carcinoma[J]. Oncol Lett, 2017,14(2):1233-1239. DOI: 10.3892/ol.2017.6312.
[8]	Zhang D, Lee H, Haspel JA , et al. Long noncoding RNA FOXD3-AS1 regulates oxidative stress-induced apoptosis via sponging microRNA-150[J]. FASEB J, 2017,31(10):4472-4481. DOI: 10.1096/fj.201700091R.
[9]	Nong Q, Li S, Wu Y , et al. LncRNA COL1A2-AS1 inhibits the scar fibroblasts proliferation via regulating miR-21/Smad7 pathway[J]. Biochem Biophys Res Commun, 2018,495(1):319-324. DOI: 10.1016/j.bbrc.2017.11.027.
[10]	Lin Q, Guan W, Ren W , et al. MALAT1 affects ovarian cancer cell behavior and patient survival[J]. Oncol Rep, 2018,39(6):2644-2652. DOI: 10.3892/or.2018.6384.
[11]	Wang WW, Zhou XL, Song YJ , et al. Combination of long noncoding RNA MALAT1 and carcinoembryonic antigen for the diagnosis of malignant pleural effusion caused by lung cancer[J]. Onco Targets Ther, 2018,11:2333-2344. DOI: 10.2147/OTT.S157551.
[12]	韩树坤, 李忠民, 韩冰 , 等. 长链非编码RNA MALAT1 在肝癌组织中的表达及其在肝癌细胞增殖与侵袭转移中的作用[J]. 中国普通外科杂志, 2018,27(1):49-55. DOI: 10.3978/j.issn.1005-6947.2018.01.008.
[13]	宋鹏, 龙彤, 梁培育 , 等. 长链非编码RNA-MALAT1对前列腺癌细胞增殖及凋亡的影响[J]. 肿瘤防治研究, 2019,46(5):411-416. DOI: 10.3971/j.issn.1000-8578.2019.18.1198.
[14]	Liu J, Tian W, Zhang W , et al. MicroRNA-142-3p/MALAT1 inhibits lung cancer progression through repressing β-catenin expression[J]. Biomed Pharmacother, 2019,114:108847. DOI: 10.1016/j.biopha.2019.108847.
[15]	Torre LA, Trabert B, DeSantis CE et al. Ovarian cancer statistics, 2018[J]. CA Cancer J Clin, 2018,68(4):284-296. DOI: 10.3322/caac.21456.
[16]	Chu ZP, Dai J, Jia LG , et al. Increased expression of long noncoding RNA HMMR-AS1 in epithelial ovarian cancer: an independent prognostic factor[J]. Eur Rev Med Pharmacol Sci, 2018,22(23):8145-8150. DOI: 10.26355/eurrev_201812_16506.
[17]	张万华, 宋福婷 . 血清miR-139-5p在卵巢癌患者中的表达及其临床意义[J]. 国际肿瘤学杂志, 2018,45(2):96-99. DOI: 10.3760/cma.j.issn.1673-422X.2018.02.008.
[18]	化定超, 王新宇 . 微小RNA与卵巢癌[J]. 国际肿瘤学杂志, 2017,44(8):615-618. DOI: 10.3760/cma.j.issn.1673-422X.2017.08.015.
[19]	Xie H, Liao X, Chen Z , et al. LncRNA MALAT1 inhibits apoptosis and promotes invasion by antagonizing miR-125b in bladder cancer cells[J]. J Cancer, 2017,8(18):3803-3811. DOI: 10.7150/jca.21228.
[20]	Kim J, Piao HL, Kim BJ , et al. Long noncoding RNA MALAT1 suppresses breast cancer metastasis[J]. Nat Genet, 2018,50(12):1705-1715. DOI: 10.1038/s41588-018-0252-3.
[21]	Xia C, Liang S, He Z , et al. Metformin, a first-line drug for type 2 diabetes mellitus, disrupts the MALAT1/miR-142-3p sponge to decrease invasion and migration in cervical cancer cells[J]. Eur J Pharmacol, 2018,830:59-67. DOI: 10.1016/j.ejphar.2018.04.027.