Method for analyzing influences of E2F3 gene slicing on whole blood E2F3 in prostate cancer animal model

Abstract

The invention discloses a method for analyzing influences of E2F3 gene silencing on E2F3 gene slicing on whole blood E2F3 in a prostate cancer animal model. The method comprises the following steps: clarifying a relationship between E2F3 and each of grading, staging and prognosis of the prostate cancer through immunohistochemical study of the expression of E2F3 in different prostate cancer tissues; silencing E2F3 gene by constructing an AAV virus vector for E2F3, and carrying out related in-vitro function experiment; and successfully applying the AAV virus vector by establishing a nude mouse prostate cancer model to carry out in vivo experiment, and clarifying the E2F3 mRNA expression in the nude mouse whole blood before and after the E2F3 gene silencing. The foundation is laid for subsequent prostate cancer gene therapy in vivo experiment with E2F3 as a target by adopting the E2F3 gene as an efficient target for prostate cancer gene therapy in the invention; and a recombinant RNA interference AAV virus for E2F3 can inhibit the proliferation of tumor cells, can promote cell apoptosis, effectively inhibits the tumor growth of the animal model, reduces the expression of E2F3 in peripheral blood specimens, and provides the reliable theoretical and experimental foundation for tumor gene silencing therapy with the E2F3 as the target.

Description

technical field [0001] The invention belongs to the technical field of E2F3 gene, in particular to a method for the influence of E2F3 gene silencing on whole blood E2F3 in animal model of prostate cancer. Background technique [0002] Prostate cancer is one of the most common male malignant tumors in Europe and the United States. At present, the incidence of prostate cancer in the United States has surpassed that of lung cancer, becoming the first tumor that endangers men's health. In my country, with the change of lifestyle, the aging of the population and the With the improvement of diagnostic technology, the incidence of prostate cancer has also shown a continuous increase trend. In Beijing and Shanghai and other modernized cities, prostate cancer has surpassed bladder cancer and become the most common urinary system tumor. [0003] The occurrence and development of tumors are closely related to out-of-control cell cycle regulation. Abnormal cell cycle regulation prevents ...

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Problems solved by technology

[0004] With the emergence of RNAi technology, gene silencing technology can more specifically silence target mRNA on the basis of maintaining high efficiency, thereby reducing the expression of corresponding protein, because RNAi technology works by inhibiting gene expression after transcription of host cell mRNA, using RNAi Anti-tumor gene therapy with anti-tumor technology is expected to become a new breakthrough in cancer treatment. The current difficulties in the application of RNAi technology in disease diagnosis and treatment mainly focus on two aspects, namely, the effectiveness of the target and the high efficiency of siRNAs or shRNAs target transfer. , the effectiveness of the target is an important prerequisite for RNAi application research. When designing siRNAs or shRNAs, we must attach great importance to the basic research of their targeting effectiveness; especially when entering clinical trials, we should pay more attention to the effectiveness of their design In order to avoid non-specific reactions in vivo or more serious potential adverse reactions that are not yet clear, second, the high efficiency of siRNAs or shRNAs target transport includes the stability of molecules in vivo and the transport issues involved in penetrating cell membranes In the early days of RNAi, liposomes were often used to transfect plasmid vectors or siRNA directly into cells. This situation often resulted in poor gene silencing effects due to low transfection efficiency. The emergence of AAV interference vectors is a more efficient gene silence lays the groundwork

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