摘要
目的检测胸苷激酶1(TK1)在宫颈癌中的表达,并分析其对顺铂敏感性和恶性进展的影响。方法将TK1空载质粒和TK1小干扰RNA转染至培养的宫颈癌HeLa、SiHa分别记为TK1-NC组和TK1-siRNA组。通过实时荧光定量逆转录聚合酶链反应(qRT-PCR)检测各组细胞中TK1 mRNA表达水平;以细胞计数法-8(CCK-8)检测细胞的增殖情况;以原位末端标记(TUNEL)法检测细胞的凋亡情况。裸鼠成瘤实验将5 mg·kg^(-1)顺铂(cisplatin,DDP)分别与TK1-NC和TK1-siRNA皮下注射至裸鼠体内,分别记为TK1-NC+DDP组和TK1-siRNA+DDP组,检测TK1对HeLa细胞顺铂敏感性的影响。结果TK1在HaCaT、HeLa及SiHa细胞中的表达分别为1.05±0.13,3.64±0.34,5.25±0.50。TK1-NC组HeLa、SiHa细胞中TK1的表达分别为1.67±0.18,1.81±0.17,凋亡率分别为(5.26±1.32)%,(5.22±1.34)%;TK1-siRNA组HeLa、SiHa细胞中TK1的表达分别为0.45±0.05,0.39±0.04,凋亡率分别为(36.65±3.64)%,(34.25±3.42)%。TK1-siRNA组第1、7和14天肿瘤生长体积分别为(105.30±10.20),(421.00±42.31)和(974.27±96.73)mm^(3),TK1-NC组第1、7和14天肿瘤生长体积分别为(109.21±10.12),(527.52±50.15)和(723.10±71.54)mm^(3),TK1-NC+DDP组第15、21和28天肿瘤生长体积分别为(954.34±95.87),(668.34±66.44)和(437.33±48.45)mm^(3),TK1-siRNA+DDP组第15、21和28天肿瘤生长体积分别为(947.11±94.55),(514.33±51.57)和(205.41±21.64)mm^(3)。HeLa、SiHa细胞与HaCaT细胞比较,TK1-siRNA组与TK1-NC组比较,TK1-siRNA+DDP组与TK1-NC+DDP组比较,差异均有统计学意义(均P<0.05)。沉默宫颈癌细胞中TK1表达后,细胞增殖和存活率显著降低,肿瘤细胞对顺铂敏感性增强。体内成瘤实验显示结果沉默TK1后裸鼠肿瘤体积显著缩小,生长速度减慢,对顺铂敏感性增加。结论沉默TK1可以抑制宫颈癌细胞的增殖,促进细胞凋亡,并且能够增强宫颈癌细胞对顺铂敏感性。
Objective To detect the expression of thymidine kinase 1(TK1)in cervical cancer and analyze its influence on cisplatin(DDP)sensitivity and malignant progression.Methods The TK1 empty plasmid and TK1 small interfering RNA were transfected into the cultured cervical cancer HeLa and SiHa as TK1-NC group and TK1-siRNA group,respectively.Real-time quantitative polymerase chain reaction(qRT-PCR)was used to detect the TK1 mRNA expression level in each group of cells;cell counting kit-8(CCK-8)method was used to detect cell proliferation;in situ end labeling(TUNEL)method was used to detect cell apoptosis.In nude mice tumor formation experiment,5 mg·kg^(-1)DDP with TK1-NC or TK1-siRNA was injected subcutaneously into nude mice,recorded as TK1-NC+DDP group and TK1-siRNA+DDP group,respectively.The effect of TK1 on the sensitivity of HeLa cells to DDP was detected.Results The expression of TK1 in HaCaT,HeLa and SiHa cells were 1.05±0.13,3.64±0.34,5.25±0.50,respectively.After transfection,the expression of TK1-NC group in HeLa and SiHa cells were 1.67±0.18 and 1.81±0.17,respectively,and the apoptosis rate were(5.26±1.32)%and(5.22±1.34)%respectively;the expression of TK1 in TK1-siRNA group in HeLa and SiHa cells were 0.45±0.05 and 0.39±0.04,and the apoptosis rate were(36.65±3.64)%and(34.25±3.42)%,respectively.The tumor growth volumes on the 1,7 and 14 days in TK1-siRNA group were(105.30±10.20),(421.00±42.31)and(974.27±96.73)mm^(3),respectively.The tumor growth volumes in TK1-NC group on the 1,7 and 14 days were(109.21±10.12),(527.52±50.15)and(723.10±71.54)mm^(3),respectively.The tumor growth volume on the 15,21 and 28 days in TK1-NC+DDP group were(954.34±95.87),(668.34±66.44)and(437.33±48.45)mm^(3).The tumor growth volume on days 15,21 and 28 in TK1-siRNA+DDP group were(947.11±94.55),(514.33±51.57)and(205.41±21.64)mm^(3),respectively.HeLa and SiHa cells compared with HaCaT cells,TK1-siRNA group compared with TK1-NC group,TK1-siRNA+DDP group compared with TK1-NC+DDP group,the difference were all statistic
作者
李琼珍
王嫱
俞海芬
李丹丹
任冬燕
LI Qiong-zhen;WANG Qiang;YU Hai-fen;LI Dan-dan;REN Dong-yan(Health Management Center,Shizong People’s Hospital,Qujing 655700,Yunnan Province,China;Department of Gynaecology,Shizong People’s Hospital,Qujing 655700,Yunnan Province,China;Cadre Physical Examination Center,Affiliated Hospital of Yunnan University,Kunming 650021,Yunnan Province,China;Department of Clinical Psychology,Yunnan First People’s Hospital,Kunming 650034,Yunnan Province,China;Department of Gynaecology,Yunnan First People’s Hospital,Kunming 650034,Yunnan Province,China)
出处
《中国临床药理学杂志》
CAS
CSCD
北大核心
2021年第20期2777-2779,2801,共4页
The Chinese Journal of Clinical Pharmacology
关键词
宫颈癌
胸苷激酶1
顺铂
增殖
凋亡
cervical cancer
thymidine kinase 1
cisplatin
proliferation
apoptosis
