吉林大学学报(医学版) ›› 2024, Vol. 50 ›› Issue (5): 1330-1338.doi: 10.13481/j.1671-587X.20240517

• 基础研究 • 上一篇    

5-Aza-CdR对裸鼠皮下移植瘤组织中甲状腺乳头状癌细胞自噬和凋亡的影响及其机制

石玉宵,刘美岚,朱美霖,魏枫()   

  1. 内蒙古科技大学包头医学院第一附属医院内分泌科,内蒙古 包头 014010
  • 收稿日期:2023-11-29 出版日期:2024-09-28 发布日期:2024-10-28
  • 通讯作者: 魏枫 E-mail:1135172562@qq.com
  • 作者简介:石玉宵(1995-),女,黑龙江齐齐哈尔市人,在读硕士研究生,主要从事甲状腺肿瘤发病机制方面的研究。
  • 基金资助:
    内蒙古自治区科技厅科技计划项目(201802128)

Effects of 5-Aza-CdR on autophagy and apoptosis of papillary thyroid cancer cells in subcutaneous xenograft tumor tissue of nude mice and its mechanism

Yuxiao SHI,Meilan LIU,Meilin ZHU,Feng WEI()   

  1. Department of Endocrinology,First Affiliated Hospital,Baotou Medical College,Inner Mongolia University of Science and Technology,Baotou 014010,China
  • Received:2023-11-29 Online:2024-09-28 Published:2024-10-28
  • Contact: Feng WEI E-mail:1135172562@qq.com

摘要:

目的 探讨5-氮杂-2'-脱氧胞苷(5-Aza-CdR)对裸鼠皮下移植瘤组织中TPC-1细胞自噬和凋亡的影响,并阐明其作用机制。 方法 16只BALB/c雌性裸鼠右前腋下接种人甲状腺乳头状癌(PTC)TPC-1细胞建立移植瘤模型。成瘤后裸鼠随机分为对照组和实验组,每组8只,对照组裸鼠腹腔注射生理盐水,实验组裸鼠腹腔注射5-Aza-CdR,隔日给药1次,连续给药4周。观察2组裸鼠移植瘤生长情况,末次给药后处死裸鼠,称瘤体质量。HE染色观察2组裸鼠移植瘤组织病理形态表现,免疫组织化学染色检测2组裸鼠移植瘤组织中微管相关蛋白轻链3(LC3)、B细胞淋巴瘤-2(Bcl-2)和Bcl-2相关X蛋白(Bax)蛋白表达水平,实时荧光定量PCR(RT-qPCR)法和Western blotting法检测2组裸鼠移植瘤组织中LC3、Beclin-1、Bcl-2、Bax、细胞外信号调节激酶激酶(MEK)、细胞外信号调节激酶1(ERK1)、细胞外信号调节激酶2(ERK2)、磷酸化ERK1(p-ERK1)、磷酸化ERK2(p-ERK2)mRNA和蛋白表达水平,并计算Bax/Bcl-2比值。 结果 与对照组比较,实验组裸鼠移植瘤体积和质量明显降低(P<0.01)。对照组裸鼠癌细胞数量多,排列紧密,细胞形状不规则,细胞核染色清晰,细胞核大,细胞有重叠和分叶,可见明显的病理性核分裂象,符合PTC病理特点;实验组裸鼠癌细胞数目明显减少,排列稀疏,细胞核固缩且胞核不明显,结缔组织明显增多。与对照组比较,实验组裸鼠移植瘤组织中LC3B和Beclin-1 mRNA及蛋白表达水平明显升高(P<0.05),Bax/Bcl-2比值升高(P<0.05或P<0.01),MEK、ERK1/2及p-ERK1/2 mRNA和蛋白表达水平明显降低(P<0.05或P<0.01)。 结论 5-Aza-CdR可抑制裸鼠皮下移植瘤组织中TPC-1细胞生长,诱导移植瘤细胞自噬并促进细胞凋亡,其机制可能与丝裂原活化蛋白激酶(MAPK)/MEK/细胞外信号调节激酶(ERK)信号通路抑制有关。

关键词: 甲状腺乳头状癌, 细胞自噬, 细胞凋亡, 移植瘤, 5-氮杂-2'-脱氧胞苷

Abstract:

Objective To discuss the effects of 5-aza-2'-deoxycytidine(5-Aza-CdR) on autophagy and apoptosis of the TPC-1 cells in subcutaneous xenograft tumor tissue of the nude mice, and to clarify its mechanism. Methods Sixteen female BALB/c nude mice were inoculated with human papillary thyroid carcinoma (PTC) TPC-1 cells in the right axilla to establish the xenograft tumor model. After tumor formation, the mice were randomly divided into control group and experiment group(n=8). The nude mice in control group were given an intraperitoneal injection of saline, while the nude mice in experiment group were given the intraperitoneal injection of 5-Aza-CdR, administered once every other day for four weeks. The growth status of xenograft tumor of the mice in both groups was observed, and the mice were sacrificed after the final administration and the tumor weights of the nude mice in two groups were detected. HE staining was used to observe the pathomorphology of xenograft tumor tissue of the nude mice in both groups;immunohistochemistry was used to detect the expression levels of microtubule-associated protein light chain 3 (LC3), B-cell lymphoma 2(Bcl-2), and Bcl-2-associated X protein(Bax) in xenograft tumor tissue of the nude mice in two groups;real-time fluorescence quantitative PCR(RT-qPCR) and Western blotting methods were used to detect the expression levels of LC3, Beclin-1, Bcl-2, Bax, mitogen-activated protein kinase kinase(MEK), extracellular signal-regulated kinase 1(ERK1), extracellular signal-regulated kinase 2(ERK2), phosphorylated EPK1(p-ERK1), and phosphorylated EPK2(p-ERK2) mRNA and proteins in xenograft tumor tissue of the nude mice in two groups. Results Compared with control group, the tumor volume and weight of the nude mice in experiment group were significantly decreased(P<0.01). The number of cancer cells of the nude mice in control group was high, and the cells were densely arranged, with irregular shapes, clear nuclear staining, large overlapping nuclei, and lobulation, showing significant pathological mitotic figures consistent with PTC pathological characteristics. The number of cancer cells of the nude mice in experiment group showed a significant decresing trend, and the cells were sparse arrangement, nuclear shrinkage, and less distinct nuclei, with a significant increase in connective tissue. Compared with control group, the expression levels of LC3B and Beclin-1 mRNA and proteins in xenograft tumor tissue of the nude mice in experiment group were significantly increased(P<0.05) and the ratio of and Bax/Bcl-2 was increased(P<0.05 or P<0.01), while the expression levels of MEK, ERK1/2, and p-ERK1/2 mRNA and proteins were significantly decreased(P<0.05 or P<0.01). Conclusion 5-Aza-CdR can inhibit the growth of TPC-1 cells in subcutaneous xenograft tumor tissue of the nude mice, induce the autophagy, and promote the apoptosis of the tumor cells. The mechanism may be related to the inhibition of the mitogen-activated protein kinase(MAPK)/MEK/extracellular signal-regulated kinase(ERK) signaling pathways.

Key words: Papillary thyroid carcinoma, Cell autophagy, Apoptosis, Transplantation tumor, 5-aza-2'-deoxycytidine

中图分类号: 

  • R736.1