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陈彩萍,徐琦.姜黄素对miR-29-VGEF的调控作用及对肝癌发生发展的影响[J].浙江中西医结合杂志,2020,30(10):
姜黄素对miR-29-VGEF的调控作用及对肝癌发生发展的影响
The regulation of curcumin on miR-29-VGEF and its effect on the development of liver cancer
投稿时间:2019-09-23  修订日期:2020-09-03
DOI:
中文关键词:  姜黄素  miR-29  VEGF  增殖  凋亡
英文关键词:
基金项目:
作者单位E-mail
陈彩萍 杭州市肿瘤医院 15088663261@189.cn 
徐琦   
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中文摘要:
      目的 研究肝癌细胞内miR-29-VEGF的表达情况和对肝癌细胞生物学过程的影响,并探索姜黄素对miR-29-VEGF的调控作用。方法 收集2008年12月1日-2018年12月1日期间就诊于我院临床资料较完整的收储切除肝癌标本41例,含癌组织和癌旁的非癌组织,采用免疫组化研究VEGF的表达情况以及与肝癌分型的相关性;转染VEGF siRNA后,MTT检测其对细胞增殖的调控作用,Western Blot 检测其对凋亡蛋白Bcl2、Bax表达的影响;PCR检测肝癌细胞内miR-29的表达,转染miR-20mimics后,qRT-PCT和Western blot 检测对VEGF的影响;MTT检测姜黄素对HepG2细胞的IC50,qRT-PCT和Western blot 检测姜黄素对miR-29-VEGF的调控作用。结果 肝癌组织中VEGF阳性率显著升高,为70.73%(29/41),且VEGF在正常肝组织、癌旁肝组织和肝癌组织中的表达呈现递增趋势,肝癌组织中的VEGF显著高于癌旁肝组织和正常肝组织,差异有显著性(p<0.001),肝癌组织中VEGF表达水平与肿瘤分化程度有关,病理分级3-4级明显高于1-2级(p<0.05);VEGF表达水平与肝癌侵袭转移性有关,高侵袭转移性有关,高侵袭转移组明显高于低侵袭转移组(p<0.05);而与肿瘤大小、血清AFP水平及HBsAg状态无相关(p>0.05);转染VEGF siRNA后,细胞的增殖受到明显抑制,细胞抗凋亡蛋白Bcl2表达显著下降,细胞凋亡蛋白Bax表达显著上升;与阴性对照组相比,抑制VEGF的表达后,HepG2细胞迁移能力明显降低;qRT-PCR检测结果显示在HepG2细胞中,miR-29的表达异常降低;转染miR-29 mimics促进miR-29的表达后,VEGF mRNA水平和蛋白质水平均显著降低;姜黄素对HepG2细胞的抑制率在72h最好,IC50为49.50umol/L;在肝癌HepG2细胞内添加姜黄素59.68umol/L作用72h后,miR-29的表达显著升高,同时VEGF表达下降。结论在肝癌细胞内,姜黄素可在某种程度上促进miR-29的表达,进而抑制VEGF的表达,调控肝癌细胞生物学过程。
英文摘要:
      objective to investigate the expression of mir-29-vegf in liver cancer cells and its effect on the biological process of liver cancer cells, and to explore the regulatory effect of curcumin on mir-29-vegf. Methods 41 cases of liver cancer specimens, including cancerous tissues and adjacent non-cancerous tissues, were collected from our hospital from December 1, 2008 to December 1, 2018. Immunohistochemistry was used to study the expression of VEGF and its correlation with liver cancer classification. After transfection with VEGF siRNA, its regulatory effect on cell proliferation was detected by MTT, and its effect on the expression of apoptotic proteins Bcl2 and Bax was detected by Western Blot. The expression of mir-29 in liver cancer cells was detected by PCR. After transfection with mir-20mimics, the effect of qrt-pct and Western blot on VEGF was detected. The effect of curcumin on mir-29-vegf was detected by MTT, qrt-pct and Western blot. Results VEGF positive rate was significantly increased in liver tissue, 70.73% (29/41), and VEGF in normal liver tissue and liver tissue and liver tissue adjacent to carcinoma in the expression showed a trend of increasing, cancer of the liver tissue of VEGF was significantly higher than that of liver tissue and normal liver tissue adjacent to carcinoma, with significant difference (p < 0.001), level of VEGF expression in liver cancer tissue associated with tumor differentiation, pathologic stage 3 or 4 level significantly higher than 1 to 2 level (p < 0.05); The expression level of VEGF was related to the invasion and metastasis of liver cancer, which was significantly higher in the high invasion and metastasis group than in the low invasion and metastasis group (p<0.05). There was no correlation with tumor size, serum AFP level or HBsAg status (p>0.05). After transfection with VEGF siRNA, cell proliferation was significantly inhibited, the expression of anti-apoptotic protein Bcl2 was significantly decreased, and the expression of apoptotic protein Bax was significantly increased. Compared with the negative control group, the migration ability of HepG2 cells was significantly reduced after inhibiting the expression of VEGF. Qrt-pcr results showed that the expression of mir-29 was abnormally reduced in HepG2 cells. After transfection with mir-29 mimics to promote the expression of mir-29, VEGF mRNA and protein levels were significantly reduced. The inhibition rate of curcumin on HepG2 cells was the best at 72h, and the IC50 was 49.50umol/L. After the addition of curcumin in HCC HepG2 cells for 72h at 59.68umol/L, the expression of mir-29 was significantly increased, while the expression of VEGF was decreased. Conclusion curcumin can promote the expression of mir-29, inhibit the expression of VEGF and regulate the biological process of liver cancer cells to some extent.
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