西红花提取物调控免疫细胞,提高程序性死亡受体-1抑制剂治疗肺腺癌效果的实验研究

Experimental study of saffron extracts to modulate immune cells to improve the efficacy of a programmed death-1 inhibitor in the treatment of lung adenocarcinoma

目的:多项研究提示,西红花提取物能影响肿瘤的发展进程。本实验探究西红花提取物在肺腺癌小鼠模型中对肿瘤免疫微环境和免疫治疗的影响,为西红花提取物抗肿瘤研究提供更多基础性数据。方法:构建Lewis肺癌细胞和萤光素酶稳定结合的小鼠皮下瘤模型,观察西红花提取物对小鼠皮下瘤和肿瘤免疫微环境的影响:运用活体成像技术跟踪肿瘤生长情况;运用流式细胞技术检测小鼠CD4^(+)、CD8^(+)T细胞的数量及占比;运用反转录-聚合酶链式反应技术检测程序性死亡受体配体1、含有T细胞免疫球蛋白和黏蛋白结构域的蛋白3(T cell immunoglobulin and mucin domaincontaining protein 3,TIM3)、淋巴细胞活化基因-3(lymphocyte-activation gene-3,LAG3)、具有免疫球蛋白和ITIM结构域的T细胞免疫受体(T cell immunoreceptor with immunoglobulin and ITIM domain,TIGIT)、胸腺细胞选择相关的高迁移率族蛋白(thymocyte selection-associated high mobility group box,TOX)1、TOX2、TOX3基因的mRNA表达情况。结果:与对照组相比,给予西红花提取物能一定程度地抑制小鼠皮下瘤的生长(P<0.05),且小鼠CD4^(+)、CD8^(+)T细胞的数量及占比均增加(P<0.05),TIM3、LAG3、TIGIT、TOX1、TOX2、TOX3的基因表达均上调(P<0.05)。结论:西红花提取物能提高肺腺癌免疫微环境中的CD4^(+)、CD8^(+)T细胞的占比,增强免疫治疗的抗肿瘤作用,进而提高肺癌免疫治疗效果,抑制肺癌发展。

Objective:A number of studies have shown that saffron extracts can affect the development of tumor.This study explored the effect of saffron extract on tumor immune microenvironment and immunotherapy in a mouse model of lung adenocarcinoma so as to provide more basic data for the anti-tumor research of saffron extracts.Methods:The transplanted tumor model of Lewis lung carcinoma-luciferase in mice was established to detect the effect of saffron extracts on the transplanted tumor in vivo.At the same time,the tumor growth was tracked by in vivo imaging technique.The number and proportion of CD4^(+)and CD8^(+) T cells were determined by flow cytometry.The mRNA levels of programmed death-ligand 1,T cell immunoglobulin and mucin domain-containing protein 3(TIM3),lymphocyte-activation gene-3(LAG3),T cell immunoreceptor with immunoglobulin and ITIM domain(TIGIT),thymocyte selection-associated high mobility group box(TOX)1,TOX2 and TOX3 were detected by reverse transcription-polymerase chain reaction(RT-PCR)and immunohistochemical techniques to verify the effect of saffron extracts on the regulation of tumor immune microenvironment.Results:Compared with the control group,the administration of saffron extracts could inhibit the growth of subcutaneous tumor in mice to a certain extent,and the number and proportion of CD4^(+) and CD8^(+) T cells were increased(P<0.05),and the expression of genes encoding TIM3,LAG3,TIGIT,TOX1,TOX2 and TOX3 was up-regulated(P<0.05).Conclusion:The saffron extracts can increase the proportion of CD4^(+) and CD8^(+) T cells in tumor microenvironment,enhance the anti-tumor effect of immunotherapy,thereby improving the effect of lung cancer immunotherapy and inhibiting the development of lung cancer.