CN-121970725-A - Method for constructing animal model for treating induced aging tumor and application of model

CN121970725ACN 121970725 ACN121970725 ACN 121970725ACN-121970725-A

Abstract

The invention belongs to the technical field of animal model construction, and particularly relates to a method for constructing an animal model for treating induced aging tumor and application of the animal model. The preparation method comprises the steps of mixing tumor cells with tumor aging cells stimulated by external sources, and inoculating the mixture in situ in the lung. The method can form a mixed tumor model with controllable aging load in situ in the lung of the mouse, can be used for imaging and monitoring, and can truly reflect tumor heterogeneity after treatment, so as to be used for researching the effect of aging cells in tumor microenvironment, evaluating aging cell removal strategies and the like. On one hand, TIS which is limited to tumor tissues and is caused by targeting tumor administration which can be realized by a human tumor patient but is difficult to realize in animals in clinic can be more simulated, and on the other hand, a TIS aging tumor model is constructed more accurately, so that the research complexity of medicines and pathological mechanisms based on the TIS aging tumor model is reduced, and the conclusion is more reliable.

Inventors

LIU ZHENBAO
Man Yunqi
LIU YANFEI
CHEN QIWEN
LIU YONGBO
Tan Yifu

Assignees

中南大学

Dates

Publication Date: 20260505
Application Date: 20251231

Claims (10)

1. The method for constructing the animal model for treating the induced aging tumor is characterized in that tumor cells and tumor aging cells stimulated by external sources are mixed and inoculated in situ in the lung.
2. The method of claim 1, wherein the tumor cells comprise LLC-Luc cells.
3. The method of claim 1, wherein the animal comprises a mouse.
4. The method of claim 1, wherein the ratio of the number of tumor cells to the number of tumor senescent cells is 1:1-1:3.
5. The method according to claim 1, wherein the tumor cells are induced by stimulation of tumor cells with doxorubicin hydrochloride.
6. The construction method according to claim 5, wherein, The tumor cells are planted in a porous plate, the culture medium is discarded after the tumor cells grow in an adherence way, the culture medium containing the doxorubicin hydrochloride is added for culture, then the culture medium is discarded, the culture medium containing the doxorubicin hydrochloride is added for continuous culture, and then the culture medium is replaced for culture.
7. The construction method according to claim 6, wherein, Cells were planted in a multi-well plate, grown on the wall for 12 hours, the medium was discarded, a DMEM basal medium containing 1. Mu.M doxorubicin hydrochloride was added, and cultured for 24 hours, then the medium was discarded, a DMEM complete medium containing 300 nM doxorubicin was added, and further cultured for 24 hours, and then the DMEM complete medium was replaced for two days.
8. The method of claim 1, wherein the model is obtained by injecting in situ a total volume of 30-70 μl of PBS-washed PBS cell suspension into the lung, and the total cell mass is 5 x 10 5 to 1 x 10 7 , and the growth cycle is 5-8 days.
9. Use of a treatment-induced aging tumor animal model according to any one of claims 1-8 for studying treatment-induced aging tumors.
10. The use according to claim 9 for screening for drugs for the treatment of tumors or for anti-aging, or for studying tumor pathogenesis or anti-aging mechanisms.

Description

Method for constructing animal model for treating induced aging tumor and application of model Technical Field The invention belongs to the technical field of animal model construction, and particularly relates to a construction method of a treatment-induced aging tumor animal model and application of the model. Background Therapeutic cellular aging (TIS-induced senescence) during tumor therapy has been widely reported and is considered an important biological event affecting tumor recurrence, metastasis and therapeutic response. A number of reviews have shown that chemotherapy or radiation therapy induces phenotypically stable senescent cells in tumor cells and surrounding tissues that, by secreting pro-inflammatory and pro-growing SASP (senescence-associated secretory phenotype) molecules, can both inhibit tumor proliferation and promote tumor regeneration and microenvironment remodeling over a long period of time, thereby affecting efficacy and prognosis. In vivo model construction, the strategies commonly used by researchers include tumor animal models treated systemically or locally by chemotherapy/radiotherapy, inducing tumor cells to age by first seeding the tumor and then injecting a chemotherapeutic drug or D-gal intravenously or locally, and evaluating on the basis of this tumor growth, immune response and efficacy of treatment strategies against aging tumor cells (senolytics). The model can directly reflect the complex interaction of the tumor and host tissues under the induction of the drug, but also accompanies the systemic toxicity of the drug to peripheral non-tumor tissues and the wide influence on non-tumor cells, thereby increasing the complexity of result analysis. The defects include: 1. The scheme of directly using chemotherapeutic drugs to induce TIS in vivo can act on tumor cells and host normal cells simultaneously, and meanwhile, compared with human clinical treatment, the targeting drugs and induction modes of murine sources have poor targeting and large peripheral toxicity. Inflammatory effects and tissue damage caused by peripheral toxicity can interfere with aging-related studies, e.g., aging-related secretory phenotype (SASP) levels in the blood can no longer be accurate due to peripheral inflammatory effects. 2. It is difficult to distinguish the senescence effect generated by the tumor itself from the secondary effect caused by the drug toxicity of the target organ and the senescence effect of the normal cells of the target organ, thereby being unfavorable for the clear analysis of the causal relationship. For example, the peripheral drug stimulated modeling method not only can cause the aging of lung cancer cells, but also can cause the aging of normal cells in the lung, so that the TIS phenomenon of lung cancer is difficult to strip for research. In addition, there are also syngeneic or xenograft models of in vitro post-aging-induced transplantation, i.e., tumor cells or stromal cells are first induced to age in vitro with chemicals (e.g., doxorubicin/doxorubicin, etc.), and then transplanted into animals for subsequent study. The method is convenient for controlling the source and proportion of the aging cells, but the aging lung cancer cells do not have proliferation capacity and hardly have tumorigenicity. As a lung cancer model, lewis lung carcinoma (light-emitting marker strains such as LLC/LLC-Luc) is a common C57BL6 strain mouse lung cancer model cell, and lung tumors can be established by intravenous injection, thoracic cavity/in-situ inoculation and other modes, and tumor load and metastasis processes can be tracked by means of in-vivo bioluminescence. The models have wide application value in the aspects of researching lung cancer biology and medicine curative effect, but the accurate construction of animal models of TIS in-situ lung cancer has not been reported yet. In summary, the prior art still has significant blank or limitation in realizing the "in situ, controllable and monitorable construction of a model of treatment-induced coexistence of senescent cells and non-senescent tumor cells in the mouse lung". In order to fill the technical blank, the development of a model method which can control the load of senescent cells and keep in-situ growth and in-vivo dynamic monitoring has important theoretical value and application prospect, and has direct significance for evaluating the strategy of removing senescent cells (senolytic), analyzing the effect of SASP in tumor recurrence/metastasis and improving tumor treatment schemes. Disclosure of Invention Aiming at the problem that the existing mouse lung cancer model cannot stably and controllably construct a model limited by the coexistence state of treatment-induced aging (treatment-Induced Senescence, TIS) tumor cells and non-aging tumor cells occurring in lung cancer tumors, the invention aims to provide the application of the model constructed by the novel animal model construction method. The method can form