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

Abstract

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 a 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 wherein the reagent for detecting the expression level of PRR11 protein includes a PCR primer capable of specifically amplifying the mRNA of PRR11 gene; further discloses the application of PRR11 gene in the preparation of drugs for treating RB tumors; further discloses a RB detection kit and a biochip containing the RB diagnostic reagent. 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

Technical Field

[0001] The present invention relates to the technical field of retinoblastoma therapeutic drugs, and more specifically, to the application of PRR11 gene in early diagnosis, prognosis evaluation and targeted treatment of retinoblastoma. Background Art

[0002] Retinoblastoma (RB) is a primary intraocular malignant tumor that occurs frequently in infants and young children, with a global incidence of approximately 1 / 15,000 to 1 / 20,000. RB can cause retinal detachment and necrosis, and invade the optic nerve and central nervous system, with a high blindness rate, which seriously threatens the quality of life of children. Late-stage RB patients can metastasize to the brain and distant sites through the optic nerve or choroid, endangering their lives. There are approximately 1,100 new patients in my country each year, most of whom are late-stage high-risk patients.

[0003] Although there are many treatments for RB, including surgery, radiotherapy, chemotherapy, laser and cryotherapy, these treatments still have some shortcomings. First, radiotherapy and chemotherapy can cause serious side effects and long-term complications; second, the treatment of RB, especially for high-risk patients, still faces a high risk of recurrence; finally, current treatment plans often adopt a "one-size-fits-all" approach and lack individualized treatment for individual patients' conditions and genetic characteristics. Biallelic mutations in the tumor suppressor gene RB1 (Retinoblastoma 1) are the initial events of malignant transformation of RB, but the continued growth and spread of tumors require additional genetic changes. At present, there is a lack of clinical biomolecular markers that can accurately assess the progression of RB and monitor recurrence. At the same time, although some molecular mechanism-based targeted therapies have made progress in the treatment of other types of cancer, the development of targeted therapeutic drugs for RB is still lagging behind, which means that treatment options are limited and the latest scientific research results are not fully utilized.

[0004] PRR11 (Proline-Rich Protein 11) is a protein encoded by the PRR11 gene on human chromosomes, which is located in the 17q22-23 region. PRR11 protein plays a role in the cell cycle process, thus playing a key role in cell proliferation, differentiation and other important cell functions.

[0005] However, so far, there are no reports on the relationship between PRR11 and RB, no reports on evaluating the prognosis of RB by detecting the expression level of PRR11, and no reports on treating RB by reducing its expression level; therefore, the present invention aims to provide the application of PRR11 gene in early diagnosis, prognosis evaluation and targeted treatment of retinoblastoma. Summary of the invention

[0006] The purpose of the present invention is to provide an application of the PRR11 gene in the early diagnosis, prognosis evaluation and targeted treatment of retinoblastoma. By combining the detection of PRR11 gene expression and targeted treatment based on the gene, it 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 circumstances of each patient, optimize the treatment effect and reduce side effects.

[0007] The above technical purpose of the present invention is achieved through the following technical solution: a RB diagnostic reagent, which contains a reagent that can detect the expression level of PRR11 protein.

[0008] The present invention is further configured as follows: the reagent for detecting the expression level of PRR11 protein includes a polyclonal antibody or a monoclonal antibody of PRR11.

[0009] The present invention is further configured as follows: the reagent for detecting the expression level of PRR11 protein includes PCR primers that can specifically amplify PRR11 gene mRNA.

[0010] The present invention also provides a RB detection kit containing the RB diagnostic reagent.

[0011] The present invention also provides a biochip containing the RB diagnostic reagent.

[0012] The present invention also provides the use of PRR11 gene in preparing medicine for treating RB tumors.

[0013] In summary, the present invention has the following beneficial effects:

[0014] 1. The present invention provides a more accurate early diagnosis method for retinoblastoma by using the PRR11 gene as a molecular biomarker, which is used to improve the accuracy of early diagnosis of RB;

[0015] 2. The present invention can develop targeted treatment strategies based on the PRR11 gene, using specific drugs or intervention methods to directly act on tumor cells. By targeting the PRR11 gene and its signaling pathway, the growth of RB tumors can be effectively inhibited. By combining the detection of PRR11 gene expression and targeted treatment based on this gene, the present invention supports the formulation of more individualized treatment plans, which helps to adjust the treatment strategy according to the specific circumstances of each patient, optimize the treatment effect and reduce side effects. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] Figure 1 This is a comparison chart of the fold changes of genes that are specifically highly expressed in RB mined using the GEO database in an embodiment of the present invention;

[0017] Figure 2 Schematic diagram of the expression of the specifically highly expressed genes in the invasive RB tissue and the non-invasive RB tissue in the embodiment of the present invention;

[0018] Figure 3 It is a comparison diagram of the protein expression of PRR11 in human retinal pigment epithelial cells and RB tumor cells Y79 and Weri-Rb1 in the embodiment of the present invention;

[0019] Figure 4 This is a diagram showing the effect of expressing PRR11 protein on the proliferation ability of Y79 and Weri-Rb1 cells in an example of the present invention. DETAILED DESCRIPTION

[0020] The following is combined with Figure 1-4 The present invention is described in further detail.

[0021] Embodiment: Application of PRR11 gene in early diagnosis, prognosis assessment and targeted treatment of retinoblastoma, including providing a RB diagnostic reagent, which contains a reagent capable of detecting the expression level of PRR11 protein, wherein the reagent for detecting the expression level of PRR11 protein comprises a polyclonal antibody or a monoclonal antibody of PRR11, and the reagent for detecting the expression level of PRR11 protein comprises a PCR primer capable of specifically amplifying PRR11 gene mRNA; the present invention also provides a RB detection kit, which contains the above-mentioned RB diagnostic reagent; the present invention also provides a biochip, which contains the above-mentioned RB diagnostic reagent.

[0022] This embodiment develops a targeted therapy strategy based on the PRR11 gene as a target. This strategy includes, but is not limited to, the use of small molecule inhibitors, monoclonal antibodies, siRNA or CRISPR-Cas9 gene editing technology to directly target and regulate the expression or function of the PRR11 gene. By inhibiting the overexpression of PRR11 or its induced signaling pathway, the proliferation of tumor cells can be effectively inhibited, thereby achieving the purpose of treating retinoblastoma.

[0023] This embodiment provides a method for screening a drug for treating RB tumors, the method comprising the following steps:

[0024] 1. Establish a cell model or animal model group of RB and a normal control group;

[0025] 2. Use the drug to be screened in a dose gradient to act on the model group, and then detect the protein expression level of PRR11 or the mRNA content of its gene;

[0026] 3. Compare the test results with those of the normal control group. If the drug to be screened can make the protein expression level of PRR11 or the mRNA content of its gene in the model group significantly approach that of the normal control group, it will be screened.

[0027] This embodiment combines the detection and evaluation results of the PRR11 gene expression level with the targeted treatment strategy to support the development of an individualized treatment plan for retinoblastoma patients. Based on the expression of the patient's PRR11 gene, doctors can choose the most appropriate treatment method, including a combination of targeted therapy and traditional treatment methods, to optimize the treatment effect, reduce side effects, and improve the patient's quality of life.

[0028] This example uses the GEO database to mine genes that are specifically highly expressed in RB. Specifically, the differentially expressed genes are mined using the screening criteria of |log2FC|>1 and P-value<0.05, such as Figure 1 As shown, (A) in the Series GSE110811 dataset, 450 differentially expressed genes were found between the RB group and the Normal retina group, of which 176 genes were up-regulated in RB tissues and 274 genes were down-regulated in RB tissues; (B) in the Series GSE24673 dataset, 1217 differentially expressed genes were found between the RB group and the Normal retina group, of which 437 genes were up-regulated in RB tissues and 780 genes were down-regulated in RB tissues; (C) in the Series GSE97508 dataset, non-invasive RB and invasive RB were both considered as RB disease groups, and adult retina was considered as the normal control group. A total of 2570 differentially expressed genes were found between the RB group and the Normal retina group, of which 1036 genes were up-regulated in RB tissues and 1534 genes were down-regulated in RB tissues; (D) in the Series GSE97508 dataset, non-invasive RB and invasive RB were considered as the RB disease groups, and adult retina was considered as the normal control group. RB was regarded as the control group. A total of 9558 differentially expressed genes were found in the invasive RB group compared with the control group, of which 5206 genes were up-regulated in invasive RB tissues and 4352 genes were down-regulated in invasive RB tissues.

[0029] Mining genes that are specifically highly expressed in RB using the GEO database revealed that the PRR11 gene was significantly highly expressed in RB compared with normal retinal tissue, and more importantly, the PRR11 gene was more highly expressed in the invasive RB group compared with the non-invasive RB group.

[0030] In this example, the GEO database was used to mine genes that were highly expressed in RB, such as Figure 2It can be concluded that only three genes (MCM3, MKI67, PRR11) are upregulated in RB tissues, and their expression is increased in invasive RB tissues compared with non-invasive RB tissues. Among them, MCM3 and MKI67 are star molecules, but the molecular function and regulatory mechanism of PRR11 in the occurrence, development and metastasis of RB are still unclear. So far, there are no reports on the relationship between PRR11 and RB, no reports on evaluating the prognosis of RB by detecting the expression level of PRR11, and no reports on treating RB by reducing its expression level.

[0031] In this example, the GEO database was used to mine genes that were highly expressed in RB, such as Figure 3 The results show that the protein expression of PRR11 was significantly increased in RB tumor cells Y79 and Weri-Rb1 compared with human retinal pigment epithelial cells (isolated from normal human retinal tissue).

[0032] In this example, the GEO database was used to mine genes that were highly expressed in RB, such as Figure 4 It can be concluded that after expressing PRR11 protein (A, B), the proliferation ability of Y79 and Weri-Rb1 cells was significantly enhanced (C, D). After knocking down PRR11 protein using the shRNA sequence of PRR11 (5'-ACTTCAGGCTGGACCATTAAA-3'), the proliferation ability of Y79 and Weri-Rb1 cells was significantly weakened (C, D).

[0033] This specific embodiment is merely an explanation of the present invention and is not a limitation of the present invention. After reading this specification, those skilled in the art may make non-creative modifications to the present embodiment as needed. However, as long as they are within the scope of the claims of the present invention, they are protected by the patent law.

Claims

1. Application of reagents for detecting the expression level of PRR11 protein in the preparation of diagnostic reagents for retinoblastoma.

2. Use of the reagent for detecting the expression level of PRR11 protein according to claim 1 in the preparation of a diagnostic reagent for retinoblastoma, characterized in that: The reagent for detecting the expression level of PRR11 protein includes a polyclonal antibody or a monoclonal antibody of PRR11.

3. Use of the reagent for detecting the expression level of PRR11 protein according to claim 1 in the preparation of a diagnostic reagent for retinoblastoma, characterized in that: The reagent for detecting the expression level of PRR11 protein includes PCR primers that can specifically amplify PRR11 gene mRNA.

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