miR-19a-3p通过PTEN调节人胃癌细胞增殖水平
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摘要
目的探讨miR-19a-3p通过下游靶基因PTEN调节人胃癌MKN28细胞增殖水平,从microRNA角度阐明胃癌细胞增殖的机制,为胃癌的防治提供新的思路。方法收集20例人胃癌组织和癌旁组织,实时荧光定量PCR方法分析miR-19a-3p表达水平;在人胃癌MKN28细胞中转染miR-19a-3p类似物和miR-19a-3p反义链,用实时荧光定量PCR方法分析miR-19a-3p表达水平,用MTT法检测细胞增殖水平;在人胃癌MKN28细胞中转染miR-19a-3p类似物和miR-19a-3p反义链,用Western blot方法分析PTEN蛋白水平;在人胃癌MKN28细胞中共转染miR-19a-3p反义链和PTEN特异性的小RNA片段(siRNA-PTEN,si-PTEN),用Western blot方法分析PTEN蛋白水平,用MTT法检测细胞增殖水平。结果胃癌组织中miR-19a-3p表达水平明显高于癌旁组织(P<0.05)。转染miR-19a-3p类似物组MKN28细胞中miR-19a-3p水平和细胞增殖水平均明显高于阴性对照组(P<0.05),转染miR-19a-3p反义链组MKN28细胞中miR-19a-3p水平和细胞增殖水平均明显低于反义阴性对照组(P<0.05)。转染miR-19a-3p类似物组与阴性对照组PTEN mRNA水平差异无统计学意义(P>0.05),miR-19a-3p反义链组与反义阴性对照组PTEN mRNA水平差异无统计学意义(P>0.05)。miR-19a-3p反义链组MKN28细胞增殖水平低于反义阴性对照组,PTEN siRNA组MKN28细胞增殖水平高于反义阴性对照组,miR-19a-3p反义链+PTEN siRNAMKN28细胞增殖水平低于PTEN siRNA组(P<0.05)。结论在人胃癌MKN28细胞中miR-19a-3p通过调节下游靶基因PTEN影响细胞增殖。
Objective To discuss the effect of miR-19a-3p on the proliferation of human gastric cancer cell via targeting PTEN, which provided new ideas for the prevention and treatment of gastric cancer. Methods Gastric carcinoma tissue specimens and matched adjacent tissues were obtained from twenty patients who were identified as aggressive gastric adenocarcinoma. Real-time PCR was used to analyze the levels of miR-19a-3p in gastric carcinoma tissue specimens and matched adjacent tissues. miR-19a-3p mimic and miR-19a-3p inhibitor were transfected into MKN28 cells. Real-time PCR was used to analyze the level of miR-19a-3p. And MTT assay was used to determine the proliferation of MKN28 cell. miR-19a-3p mimic and miR-19a-3p inhibitor were transfected into MKN28 cells. Western blot was used to measure the protein level of PTEN. miR-19a-3p inhibitor and siRNA-PTEN were transfected into MKN28 cells. Western blot was used to measure the protein level of PTEN. And MTT assay was used to determine the proliferation of MKN28 cell. Results Compared with adjacent tissues, miR-19a-3p expression was increased in gastric cancer tissues(P<0.05). Compared with negative mimic control group, transfection with miR-19a-3p mimic increased miR-19a-3p expression(P<0.05). Compared with negative inhibitor control group, transfection with miR-19a-3p inhibitor decreased miR-19a-3p expression(P<0.05). The level of PTEN mRNA did not significantly change in MKN28 cells transfected with miR-19a-3p mimic(P>0.05) or miR-19a-3p inhibitor(P>0.05). The proliferation of MKN28 cells was inhibited in MKN28 cells transfected with miR-19a-3p inhibitor(P<0.05) and was promoted in MKN28 cells transfected with si-PTEN(P<0.05). Compared with si-PTEN group, the proliferation of MKN28 cells was decreased in MKN28 cells co-transfected with si-PTEN and miR-19a-3p inhibitor(P<0.05). Conclusion miR-19a-3p regulated the proliferation of human gastric cancer cells via targeting PTEN.
Objective To discuss the effect of miR-19a-3p on the proliferation of human gastric cancer cell via targeting PTEN, which provided new ideas for the prevention and treatment of gastric cancer. Methods Gastric carcinoma tissue specimens and matched adjacent tissues were obtained from twenty patients who were identified as aggressive gastric adenocarcinoma. Real-time PCR was used to analyze the levels of miR-19a-3p in gastric carcinoma tissue specimens and matched adjacent tissues. miR-19a-3p mimic and miR-19a-3p inhibitor were transfected into MKN28 cells. Real-time PCR was used to analyze the level of miR-19a-3p. And MTT assay was used to determine the proliferation of MKN28 cell. miR-19a-3p mimic and miR-19a-3p inhibitor were transfected into MKN28 cells. Western blot was used to measure the protein level of PTEN. miR-19a-3p inhibitor and siRNA-PTEN were transfected into MKN28 cells. Western blot was used to measure the protein level of PTEN. And MTT assay was used to determine the proliferation of MKN28 cell. Results Compared with adjacent tissues, miR-19a-3p expression was increased in gastric cancer tissues(P<0.05). Compared with negative mimic control group, transfection with miR-19a-3p mimic increased miR-19a-3p expression(P<0.05). Compared with negative inhibitor control group, transfection with miR-19a-3p inhibitor decreased miR-19a-3p expression(P<0.05). The level of PTEN mRNA did not significantly change in MKN28 cells transfected with miR-19a-3p mimic(P>0.05) or miR-19a-3p inhibitor(P>0.05). The proliferation of MKN28 cells was inhibited in MKN28 cells transfected with miR-19a-3p inhibitor(P<0.05) and was promoted in MKN28 cells transfected with si-PTEN(P<0.05). Compared with si-PTEN group, the proliferation of MKN28 cells was decreased in MKN28 cells co-transfected with si-PTEN and miR-19a-3p inhibitor(P<0.05). Conclusion miR-19a-3p regulated the proliferation of human gastric cancer cells via targeting PTEN.
引文
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[2] Hwang JH.Understanding gastric cancer risk factors:we need to close the gap[J].Gut Liver,2018,12(1):1-2.
[3] 黄耀孟,吴珺,李秉慧.miRNA在乳腺癌诊断/预后和预测中的作用[J].河北医科大学学报,2018,39(9):1100-1106.
[4] Wu N,Han Y,Liu H,et al.miR-5590-3p inhibited tumor growth in gastric cancer by targeting DDX5/AKT/m-TOR pathway[J].Biochem Biophys Res Commun,2018,503(3):1491-1497.
[5] Liu F,Hu H,Zhao J,et al.miR-124-3p acts as a potential marker and suppresses tumor growth in gastric cancer[J].Biomed Rep,2018,9(2):147-155.
[6] Hua K,Chen YT,Chen CF,et al.MicroRNA-23a/27a/24-2 cluster promotes gastric cancer cell proliferation synergistically[J].Oncol Lett,2018,16(2):2319-2325.
[7] Li S,Zhang T,Zhou X,et al.The tumor suppressor role of miR-155-5p in gastric cancer[J].Oncol Lett,2018,16(2):2709-2714.
[8] Fuziwara CS,Kimura ET.Insights into regulation of the miR-17-92 cluster of miRNAs in cancer[J].Front Med(Lausanne),2015,2:e64.
[9] Chen M,Lin M,Wang X.Overexpression of miR-19a inhibits colorectal cancer angiogenesis by suppressing KRAS expression[J].Oncol Rep,2018,39(2):619-626.
[10] Wang Y,Zhao S,Zhu L,et al.MiR-19a negatively regulated the expression of PTEN and promoted the growth of ovarian cancer cells[J].Gene,2018,670:166-173.
[11] Bai Y,Lin H,Fang Z,et al.Plasma microRNA-19a as a potential biomarker for esophageal squamous cell carcinoma diagnosis and prognosis[J].Biomark Med,2017,11(5):431-441.
[12] Nie H,Mu J,Wang J,et al.miR195-5p regulates multi-drug resistance of gastric cancer cells via targeting ZNF139[J].Oncol Rep,2018,40(3):1370-1378.
[13] Wu Q,Yang Z,An Y,et al.MiR-19a/b modulate the metastasis of gastric cancer cells by targeting the tumour suppressor MXD1[J].Cell Death Dis,2014,5:e1144.
[14] Qiao F,Gong P,Song Y,et al.Downregulated PITX1 modulated by MiR-19a-3p promotes cell malignancy and predicts a poor prognosis of gastric cancer by affecting transcriptionally activated PDCD5[J].Cell Physiol Biochem,2018,46(6):2215-2231.
[15] Lu WD,Zuo Y,Xu Z,et al.MiR-19a promotes epithelial-mesenchymal transition through PI3K/AKT pathway in gastric cancer[J].World J Gastroenterol,2015,21(15):4564-4573.
[16] Milella M,Falcone I,Conciatori F,et al.PTEN:multiple functions in human malignant tumors[J].Front Oncol,2015,5:e24.
[17] Bermúdez Brito M,Goulielmaki E,Papakonstanti EA.Focus on PTEN Regulation[J].Front Oncol,2015,5:e166.
[18] Wang X,Huang H,Young KH.The PTEN tumor suppressor gene and its role in lymphoma pathogenesis[J].Aging(Albany NY),2015,7(12):1032-1049.