CN118348247B - Application of PRR11 gene in early diagnosis, prognosis evaluation and targeted therapy of retinoblastoma - Google Patents

Application of PRR11 gene in early diagnosis, prognosis evaluation and targeted therapy of retinoblastoma Download PDF

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CN118348247B
CN118348247B CN202410637864.8A CN202410637864A CN118348247B CN 118348247 B CN118348247 B CN 118348247B CN 202410637864 A CN202410637864 A CN 202410637864A CN 118348247 B CN118348247 B CN 118348247B
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prr11
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龚波
何宇
李欢
肖嘉陵
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Sichuan Academy Of Medical Sciences Sichuan Provincial People's Hospital
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Abstract

本发明涉及视网膜母细胞瘤治疗药物技术领域，公开了PRR11基因在视网膜母细胞瘤早期诊断、预后评估与靶向治疗中的应用，包括一种RB诊断试剂，所述RB诊断试剂含有可检测PRR11蛋白表达水平的试剂，所述检测PRR11蛋白表达水平的试剂包括PRR11的多克隆抗体或单克隆抗体，所述检测PRR11蛋白表达水平的试剂包括能特异扩增PRR11基因mRNA的PCR引物；还公开了PRR11基因在制备治疗RB肿瘤药物中的应用；还公开了一种含有RB诊断试剂的RB检测试剂盒和生物芯片。本发明支持更加个体化的治疗方案的制定，解决了现有治疗方法中评估的不准确性、高复发率以及缺乏有效的靶向治疗策略的问题，有助于根据每位患者的具体情况调整治疗策略，优化治疗效果并减少副作用。

The present invention relates to the technical field of retinoblastoma therapeutic drugs, and discloses the application of PRR11 gene in early diagnosis, prognosis evaluation and targeted treatment of retinoblastoma, including an RB diagnostic reagent, wherein the RB diagnostic reagent contains a reagent capable of detecting the expression level of PRR11 protein, wherein the reagent for detecting the expression level of PRR11 protein includes a polyclonal antibody or a monoclonal antibody of PRR11, and the reagent for detecting the expression level of PRR11 protein includes a PCR primer capable of specifically amplifying the mRNA of PRR11 gene; the application of PRR11 gene in the preparation of drugs for treating RB tumors is also disclosed; and an RB detection kit and a biochip containing the RB diagnostic reagent are also disclosed. The present invention supports the formulation of more individualized treatment plans, solves the problems of inaccurate evaluation, high recurrence rate and lack of effective targeted treatment strategies in existing treatment methods, and helps to adjust the treatment strategy according to the specific situation of each patient, optimize the treatment effect and reduce side effects.

Description

Application of PRR11 gene in early diagnosis, prognosis evaluation and targeted treatment of retinoblastoma
Technical Field
The invention relates to the technical field of retinoblastoma treatment medicines, in particular to application of PRR11 gene in early diagnosis, prognosis evaluation and targeted treatment of retinoblastoma.
Background
Retinoblastoma (Retinoblastoma, RB) is a primary intraocular malignancy, which is common to infants and young children, with a global incidence of about 1/15000 to 1/20000.RB can cause retinal detachment, necrosis and invasion of the optic nerve and central nervous system, the blindness rate is high, the life quality of the sick children is seriously threatened, the patients with advanced RB can transfer into the cranium and the distance through the optic nerve or the choroid, the life is endangered, the newly increased patients in China are about 1100 people each year, and most of patients with advanced high risk are patients.
Although current methods of treating RB are varied and cover surgery, radiation therapy, chemotherapy, laser and cryotherapy, etc., these treatments still suffer from some drawbacks. Firstly, radiotherapy and chemotherapy can cause serious side effects and long-term complications, secondly, in the RB treatment process, particularly for high-risk patients, the high recurrence risk still exists, and finally, the current treatment scheme usually adopts a one-time cutting method, and the personalized treatment aiming at the illness state and the gene characteristics of individual patients is lacking. The double allelic mutation of the oncogene RB1 (Retinoblastoma 1) is the initial event for malignant transformation of RB, but additional genetic alterations are required for continued growth and spread of tumors. Currently, there is a clinical lack of biomolecular markers that can accurately assess RB progression and monitor recurrence. Meanwhile, although some molecular mechanism-based targeted therapies have advanced in other types of cancer therapies, targeted therapeutic drug development against RB still falls behind, meaning that the treatment options are limited and the latest scientific research results have not been fully utilized.
The PRR11 (Proline-Rich Protein 11) Protein is a Protein encoded by the PRR11 gene on the human chromosome, which is located in the 17q22-23 region. PRR11 protein plays a role in cell cycle progression and thus plays a key role in cell proliferation, differentiation and other important cellular functions.
However, up to now, no report on the relation between PRR11 and RB is found, no report on the evaluation of RB prognosis by detecting the expression level of PRR11 is found, and no report on the treatment of RB by reducing the expression level is found, so that the invention aims to provide the application of PRR11 gene in early diagnosis, prognosis evaluation and targeted treatment of retinoblastoma. .
Disclosure of Invention
The invention aims to provide application of PRR11 gene in early diagnosis, prognosis evaluation and targeted therapy of retinoblastoma, and by combining detection of PRR11 gene expression and targeted therapy based on the gene, the invention supports formulation of more individual therapeutic schemes, solves the problems of inaccuracy, high recurrence rate and lack of effective targeted therapeutic strategies in the existing therapeutic methods, and is beneficial to adjusting the therapeutic strategies according to the specific conditions of each patient, optimizing the therapeutic effects and reducing side effects.
The technical aim of the invention is achieved by the following technical scheme that the RB diagnostic reagent contains a reagent capable of detecting the expression level of PRR11 protein.
The invention further provides that the reagent for detecting the expression level of PRR11 protein comprises polyclonal antibody or monoclonal antibody of PRR 11.
The invention further provides that the reagent for detecting the expression level of PRR11 protein comprises PCR primers capable of specifically amplifying PRR11 gene mRNA.
The invention also provides an RB detection kit containing the RB diagnostic reagent.
The present invention also provides a biochip containing the RB diagnostic reagent.
The invention also provides application of the PRR11 gene in preparing a medicament for treating RB tumors.
In summary, the invention has the following beneficial effects:
1. the invention provides a more accurate early diagnosis method of retinoblastoma by using PRR11 gene as a molecular biomarker, which is used for improving the accuracy of early diagnosis of RB;
2. the invention can develop a target treatment strategy based on PRR11 genes, directly acts on tumor cells by using specific drugs or intervention means, can effectively inhibit RB tumor growth by targeting PRR11 genes and signal paths thereof, and supports formulation of more individual treatment schemes by combining detection of PRR11 gene expression and target treatment based on the genes, thereby being beneficial to adjusting the treatment strategy according to specific conditions of each patient, optimizing treatment effects and reducing side effects.
Drawings
FIG. 1 is a graph showing fold change of genes specifically and highly expressed in RBs by using a GEO database in the example of the present invention;
FIG. 2 is a schematic representation of the expression of a specific high-expression gene in invasive and non-invasive RB tissues according to an embodiment of the present invention;
FIG. 3 is a graph showing the protein expression of PRR11 in human retinal pigment epithelial cells and RB tumor cells Y79 and Weri-Rb1 in accordance with an embodiment of the present invention;
FIG. 4 is a graph showing the effect of expression of PRR11 protein on proliferation potency of Y79 and Weri-Rb1 cells in the examples of the present invention.
Detailed Description
The invention is described in further detail below with reference to fig. 1-4.
The embodiment of the invention provides application of PRR11 genes in early diagnosis, prognosis evaluation and targeted treatment of retinoblastoma, comprising providing an RB diagnostic reagent containing a reagent capable of detecting the expression level of PRR11 proteins, wherein the reagent for detecting the expression level of PRR11 proteins comprises a polyclonal antibody or a monoclonal antibody of PRR11, and the reagent for detecting the expression level of PRR11 proteins comprises PCR primers capable of specifically amplifying PRR11 gene mRNA, and further provides an RB detection kit containing the RB diagnostic reagent and a biochip containing the RB diagnostic reagent.
This example developed a targeted therapeutic strategy based on PRR11 gene as target. Such strategies include, but are not limited to, direct targeting and modulation of expression or function of PRR11 genes using small molecule inhibitors, monoclonal antibodies, siRNA or CRISPR-Cas9 gene editing techniques, among others. Through inhibiting the over-expression of PRR11 or the signal path induced by the over-expression, the proliferation of tumor cells can be effectively inhibited, so that the aim of treating retinoblastoma is fulfilled.
The embodiment provides a method for screening RB tumor treatment drugs, which comprises the following steps:
1. Establishing a cell model or a model group of an animal model of RB, and establishing a normal control group;
2. Acting the medicine to be screened on the model group according to a dose gradient, and then detecting the protein expression level of PRR11 or the mRNA content of the gene thereof;
3. Comparing the detection result with the normal control group, the protein expression level of PRR11 or the mRNA content of the gene thereof in the model group can obviously approach to the drug to be screened in the normal control group, namely, the drug to be screened is screened.
In this example, the present invention supports the formulation of personalized treatment regimens for retinoblastoma patients by combining the results of detection and assessment of PRR11 gene expression levels with targeted therapeutic strategies. Depending on the expression of the PRR11 gene in the patient, the physician can select the most appropriate treatment method, including combination of targeted therapy with conventional treatment methods, to optimize the therapeutic effect, reduce side effects, and improve the quality of life of the patient.
In this example, genes specifically and highly expressed in RB were mined by using GEO database. Specifically, |log2FC| >1, P-value <0.05 was used as screening criteria to mine differential genes, as shown in FIG. 1, wherein (A) in SERIES GSE110811 dataset, there were 450 differential genes found in total, 176 genes up-regulated in RB tissue compared to Normal retina, 274 genes down-regulated in RB tissue, (B) in SERIES GSE24673 dataset, there were 1217 differential genes found in total, RB tissue compared to Normal retina, 780 genes down-regulated in RB tissue, (C) in SERIES GSE97508 dataset, there were no in-variance RB and in-variance RB were considered as RB composition disease group, adult retina as normal control group, RB group compared to Normal retina, there were 2570 differential genes found in total, 1036 genes up-regulated in RB tissue, 1534 genes down-regulated in RB tissue, (D) in SERIES GSE97508 dataset, there were no in-variance as control group, in-variance RB group compared to control group, there were no in-variance gene found in 438, and there were no in-variance found in-variance in the control group, and there were no in-variance gene found in the group at 9552.
Mining genes specifically highly expressed in RBs using the GEO database can conclude that PRR11 genes are significantly highly expressed in RBs compared to normal retinal tissue and, more importantly, PRR11 genes are expressed higher in invasive RB groups compared to non-invasive RBs.
In this example, genes specifically highly expressed in RBs were mined by the GEO database, and as shown in FIG. 2, it was concluded that only three genes (MCM 3, MKI67, PRR 11) were up-regulated in RB tissues, while expression was increased in invasive RB tissues compared to non-invasive RB tissues. Among them, MCM3 and MKI67 are star molecules, but the molecular functions and regulatory mechanisms of PRR11 in the development and transfer of RB are not clear, and up to now, no report on the relationship between PRR11 and RB has been found, no report on the evaluation of RB prognosis by detecting the expression level of PRR11, nor the achievement of RB treatment by decreasing the expression level thereof has been found.
In this example, by mining genes specifically highly expressed in RB through GEO database, it can be concluded that protein expression of PRR11 was significantly increased in RB tumor cells Y79 and Weri-Rb1 as compared to human retinal pigment epithelial cells (isolated from normal human retinal tissue), as shown in FIG. 3.
In this example, genes specifically and highly expressed in RB were mined by GEO database, and as shown in FIG. 4, it can be concluded that proliferation capacity of Y79 and Weri-Rb1 cells was significantly enhanced (C, D) after expression of PRR11 protein (A, B). After knocking down the PRR11 protein using the shRNA sequence of PRR11 (5'-ACTTCAGGCTGGACCATTAAA-3'), the proliferation capacity of Y79 and Weri-Rb1 cells was significantly reduced (C, D).
The present embodiment is only for explanation of the present invention and is not to be construed as limiting the present invention, and modifications to the present embodiment, which may not creatively contribute to the present invention as required by those skilled in the art after reading the present specification, are all protected by patent laws within the scope of claims of the present invention.

Claims (3)

1. Application of a reagent for detecting PRR11 protein expression level in preparation of retinoblastoma diagnostic reagent.
2. The use of the reagent for detecting the expression level of PRR11 protein according to claim 1, wherein the reagent for detecting the expression level of PRR11 protein comprises a polyclonal antibody or a monoclonal antibody of PRR 11.
3. The use of the reagent for detecting the expression level of PRR11 protein according to claim 1, wherein the reagent for detecting the expression level of PRR11 protein comprises PCR primers capable of specifically amplifying mRNA of PRR11 gene.
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Citations (1)

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CN102558336A (en) * 2012-01-13 2012-07-11 重庆医科大学 PRR11 gene and coded protein and application thereof

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