Application of RRM2B gene or protein thereof in metastasis of hepatocellular carcinoma

A gene and protein technology, applied in the field of application of RRM2B gene or its protein in the diagnosis and treatment of liver cancer metastasis, can solve the problem of unclear mechanism of action and specific role

Active Publication Date: 2015-05-20
SHANGHAI INST OF ONCOLOGY
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, research on the specific function and mechanism of action of these two subunits is quite controversial, especia

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  • Application of RRM2B gene or protein thereof in metastasis of hepatocellular carcinoma
  • Application of RRM2B gene or protein thereof in metastasis of hepatocellular carcinoma
  • Application of RRM2B gene or protein thereof in metastasis of hepatocellular carcinoma

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Experimental program
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Effect test

Embodiment 1

[0251] Example 1 The expression level of RRM2B gene in hepatocellular carcinoma and the correlation between the expression level of RRM2B and intrahepatic metastasis of liver cancer patients were determined.

[0252] Firstly, the expression level of RRM2B protein in 236 liver cancer tissues was analyzed by immunohistochemical method, and it was found that the expression level of RRM2B protein in liver cancer was significantly lower than that in corresponding paracancerous tissues ( figure 1 A), in addition, the expression levels of RRM2B mRNA and protein in liver cancer tissues and paracancerous tissues were detected by quantitative PCR and Western blotting, and the results were consistent with the results of immunohistochemistry, that is, the expression of RRM2B mRNA and protein in liver cancer tissues was obvious lower than the corresponding paracancerous tissue ( figure 1 B and 1C). Therefore, these results indicate that the expression of RRM2B protein is reduced in liver ...

Embodiment 2

[0257] Example 2 Determination of inhibitory effect on migration, invasion and metastasis of RRM2B liver cancer cells

[0258] 2.1 In vitro experiments

[0259] Firstly, hepatoma cell lines were infected with lentivirus to establish stable RRM2B gene silencing and overexpression hepatocellular carcinoma cell lines. The results showed that RRM2B gene silencing could significantly promote the migration and invasion of SMMC-7721 and HCC-LY10 liver cancer cells ( figure 1 D); Conversely, RRM2B gene overexpression can inhibit the migration and invasion of SMMC-7721 and Huh7 cells ( figure 1 E).

[0260] 2.2 In vivo experiments

[0261] Liver orthotopic tumor transplantation experiments in nude mice demonstrated that RRM2B gene silencing promoted the ability of SMMC-7721 and HCC-LY10 liver cancer cells to metastasize in the liver and distant lung in nude mice ( figure 1 F).

[0262] Taken together, these results indicate that RRM2B can inhibit the migration, invasion and metast...

Embodiment 3

[0263] Example 3 The status of the p53 gene does not affect the relationship between the RRM2B gene and the metastasis of liver cancer cells

[0264] 3.1 Determination of the relationship between RRM2B expression and p53 in liver cancer cell lines with known p53 levels

[0265] Since RRM2B is the target gene of the wild-type p53 gene, whether the status of the p53 gene affects the effect of RRM2B on the metastasis of HCC cells was analyzed.

[0266] Because the status of the p53 gene of some liver cancer cell lines is known, for example, the p53 gene of the liver cancer cell lines MHCC-97L, MHCC-97H, MHCC-97, SNU-182 and other cell lines are mutant, like SMMC-7721 cells The p53 gene is wild type, and the p53 gene of Hep3B cells is deletion type. Therefore, the expression levels of p53 and RRM2B proteins in liver cancer cell lines were analyzed, and the results showed that the basal expression level of the basic RRM2B protein was not related to the status of the p53 gene ( f...

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Abstract

The invention discloses application of RRM2B gene or protein thereof in metastasis of hepatocellular carcinoma, and particularly provides application of detection reagents of ribonucleoside-diphosphate reductase subunit M2B (RRM2B) gene or protein thereof, as well as the RRM2B gene or protein thereof for preparing kits for diagnosing metastasis of hepatocellular carcinoma. Furthermore, the RRM2B has an inhibiting effect on metastasis of hepatocellular carcinoma, and a new approach is provided to diagnosis and treatment of metastasis of hepatocellular carcinoma.

Description

technical field [0001] 本发明涉及肿瘤诊疗领域。具体地,本发明涉及RRM2B基因或其蛋白在肝癌转移诊断和治疗中的应用。 Background technique [0002] 原发性肝癌是一种严重危害人类健康的恶性肿瘤,而包括中国在内的东亚地区是肝癌的高发区,由于其恶性程度高、预后不良,在中国原发性肝癌已经成为第二位致死性肿瘤。其中肝细胞癌(hepatocellular carcinoma,HCC)占原发性肝癌的90%以上。研究表明肝癌的发生与病毒性肝炎及摄入黄曲霉素污染的食物等等有密切关系,其发生发展与癌基因异常表达及抑癌基因失活密切相关。HCC发生发展的分子机制十分复杂,目前尚不完全清楚。因此,进一步探索研究新的基因功能改变与肝癌发生发展及其恶性特征的关系,对揭示其发生发展的精确分子机制、设计合理的治疗方案及判断预后,进一步提高肝癌的治疗水平具有重要意义。 [0003] 核苷酸是DNA和RNA的构件分子,核苷酸生物合成的紊乱广泛影响正常细胞的生理功能,最终导致正常细胞的恶性转化,由于肿瘤细胞具有无限增殖的能力,机体必须为其需要提供更多的核苷酸。因此核苷酸代谢在肿瘤的发生和发展中发挥非常重要的作用。研究表明有一系列关键的酶调节核苷酸的合成和修饰,其中核糖核苷酸还原酶(RR)就是其中的一个。 [0004] 在人体中RR由有三个亚基构成,一个大亚基(RRM1)和两个小亚基(RRM2和RRM2B)构成。每个亚基所起的作用各不相同,如RRM1被广泛认同为抑癌基因,RRM2和RRM2B则被认为通过p53的调节影响DNA修复。然而,关于这两个亚基的具体功能和作用机制的研究颇有争议,尤其是RRM2B,目前尚未明确该基因的作用机制以及在癌症中起到的具体作用。 [0005] 因此,本领域迫切需要进行对肿瘤相关基因的研究,尤其是在中国发病率很高的肝癌的相关基因,并阐明这些基因与肝癌的发生、发展的关系,并且为肝癌的诊断和治疗提供新的思路。 Contents of the invention [0006] 本发明提供了一种诊断肝癌转移的标志物RRM2B,以及RRM2B在治疗肝癌转移中的应用。 [0007] 本发明第一方面,提供了一种核糖核苷酸还原酶亚基M2B(Ribonucleoside-diphosphate...

Claims

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Application Information

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IPC IPC(8): C12Q1/68G01N33/574G01N33/573A61K48/00A61K38/44A61P35/00C12Q1/02C12N15/85
Inventor 李锦军田华李红葛超张立行
Owner SHANGHAI INST OF ONCOLOGY
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