安徽医学
安徽醫學
안휘의학
ANHUI MEDICAL JOURNAL
2015年
4期
399-402
,共4页
宫颈癌%Hela细胞%Toll样受体9%细胞凋亡%CpG-ODN
宮頸癌%Hela細胞%Toll樣受體9%細胞凋亡%CpG-ODN
궁경암%Hela세포%Toll양수체9%세포조망%CpG-ODN
Cervical cancer%Hela cells%Toll-like receptor 9%Cell apoptosis%CpG-ODN
目的：研究宫颈癌Hela细胞中Toll样受体9（TLR9）的表达情况及TLR9对宫颈癌Hela细胞抗凋亡能力的影响，寻求宫颈癌免疫治疗的新途径。方法体外培养宫颈癌Hela细胞株，应用逆转录聚合酶链反应（RT-PCR）检测Hela细胞中TLR9 mRNA的表达情况；四甲基偶氮唑盐（MTT）检测不同浓度人肿瘤坏死因子相关凋亡诱导配体（TRAIL）对Hela细胞生长抑制情况；流式细胞术（FCM）检测CpG-ODN预刺激对Hela细胞抗TRAIL诱导凋亡能力的影响。结果①TLR9 mRNA在Hela细胞中表达，应用CpG-ODN刺激24 h后，TLR9 mRNA的表达增加，与PBS对照组比较，差异具有统计学意义（P＜0．05）。②MTT比色法检测显示， TRAIL蛋白可抑制Hela细胞生长，随着TRAIL蛋白浓度增加，抑制率增加。并且计算半抑制浓度IC50（223．092±3．850）ng／mL。③流式细胞术检测显示，TLR9特异性激动剂CpG-ODN刺激Hela细胞后，TRAIL诱导细胞凋亡率为（15．32±2．46）％，与无CpG-ODN预刺激组[（43．49±2．04）％]相比较，差异具有统计学意义（P＜0．05）。结论 TLR9特异性激动剂CpG-ODN能够增强Hela细胞抵抗TRAIL诱导的细胞凋亡能力，提示TLR9在肿瘤的恶性进程中发挥作用，为宫颈癌的免疫治疗提供新的思路。
目的：研究宮頸癌Hela細胞中Toll樣受體9（TLR9）的錶達情況及TLR9對宮頸癌Hela細胞抗凋亡能力的影響，尋求宮頸癌免疫治療的新途徑。方法體外培養宮頸癌Hela細胞株，應用逆轉錄聚閤酶鏈反應（RT-PCR）檢測Hela細胞中TLR9 mRNA的錶達情況；四甲基偶氮唑鹽（MTT）檢測不同濃度人腫瘤壞死因子相關凋亡誘導配體（TRAIL）對Hela細胞生長抑製情況；流式細胞術（FCM）檢測CpG-ODN預刺激對Hela細胞抗TRAIL誘導凋亡能力的影響。結果①TLR9 mRNA在Hela細胞中錶達，應用CpG-ODN刺激24 h後，TLR9 mRNA的錶達增加，與PBS對照組比較，差異具有統計學意義（P＜0．05）。②MTT比色法檢測顯示， TRAIL蛋白可抑製Hela細胞生長，隨著TRAIL蛋白濃度增加，抑製率增加。併且計算半抑製濃度IC50（223．092±3．850）ng／mL。③流式細胞術檢測顯示，TLR9特異性激動劑CpG-ODN刺激Hela細胞後，TRAIL誘導細胞凋亡率為（15．32±2．46）％，與無CpG-ODN預刺激組[（43．49±2．04）％]相比較，差異具有統計學意義（P＜0．05）。結論 TLR9特異性激動劑CpG-ODN能夠增彊Hela細胞牴抗TRAIL誘導的細胞凋亡能力，提示TLR9在腫瘤的噁性進程中髮揮作用，為宮頸癌的免疫治療提供新的思路。
목적：연구궁경암Hela세포중Toll양수체9（TLR9）적표체정황급TLR9대궁경암Hela세포항조망능력적영향，심구궁경암면역치료적신도경。방법체외배양궁경암Hela세포주，응용역전록취합매련반응（RT-PCR）검측Hela세포중TLR9 mRNA적표체정황；사갑기우담서염（MTT）검측불동농도인종류배사인자상관조망유도배체（TRAIL）대Hela세포생장억제정황；류식세포술（FCM）검측CpG-ODN예자격대Hela세포항TRAIL유도조망능력적영향。결과①TLR9 mRNA재Hela세포중표체，응용CpG-ODN자격24 h후，TLR9 mRNA적표체증가，여PBS대조조비교，차이구유통계학의의（P＜0．05）。②MTT비색법검측현시， TRAIL단백가억제Hela세포생장，수착TRAIL단백농도증가，억제솔증가。병차계산반억제농도IC50（223．092±3．850）ng／mL。③류식세포술검측현시，TLR9특이성격동제CpG-ODN자격Hela세포후，TRAIL유도세포조망솔위（15．32±2．46）％，여무CpG-ODN예자격조[（43．49±2．04）％]상비교，차이구유통계학의의（P＜0．05）。결론 TLR9특이성격동제CpG-ODN능구증강Hela세포저항TRAIL유도적세포조망능력，제시TLR9재종류적악성진정중발휘작용，위궁경암적면역치료제공신적사로。
Objective To evaluate the expression of TLR9 in Hela cell line and the influence of TLR9 signaling pathway on apopto-sis resistance of cervical cancer cell line Hela,in order to find a new approach of immunotherapy for cervical cancer. Methods Cervical cancer Hela cell line was cultured in vitro. Reverse transcription polymerase chain reaction (RT-PCR)was used to evaluate the expression level of TLR9 mRNA in Hela cells. Methyl thiazolyl tetrazolium (MTT)assay was used to detect the inhibition rate of cell proliferation of Hela cells which was plused with tumor necrosis factor-related apoptosis-inducing ligand (TRAIL)protein at different concentrations. Flow cytome-try(FCM)was used to measure the rate of apoptosis cell induced by TRAIL protein. Results ①The TLR9 mRNA was detected in cervical cancer Hela cell line. And the expression level of TLR9 mRNA of Hela cells incubated with TLR9 agonistic CpG-ODN was significantly high-er than that in normal controls(P<0. 05).②The cell proliferation of Hela cell was inhibited by TRAIL protein and the half maximal inhibito-ry concentration of TRAIL protein was (223. 092 ± 3. 850 )ng/ml. ③The percentage of apoptotic Hela cells treated with CpG-ODN was (15. 32 ± 2. 46)%,which decreased markedly than the control group(43. 49 ± 2. 04)% without the treatment of CpG-ODN(P<0. 05). Conclusion TLR9 agonistic ligand CpG-ODN induces the resistance of Hela cells to TRAIL-induced apoptosis,which suggests TLR9 signa-ling plays an important role in tumor progression and leads to novel immunotherapy against cervical cancer.
