临床普外科电子杂志
臨床普外科電子雜誌
림상보외과전자잡지
Journal of General Surgery for Clinicians (Electronic Version)
2014年
2期
23-27
,共5页
刘同运%刘春林%李谦%战淑慧%马贵亮%毛伟征
劉同運%劉春林%李謙%戰淑慧%馬貴亮%毛偉徵
류동운%류춘림%리겸%전숙혜%마귀량%모위정
胃癌细胞%TNF-α%慢病毒%自分泌%杀伤作用
胃癌細胞%TNF-α%慢病毒%自分泌%殺傷作用
위암세포%TNF-α%만병독%자분비%살상작용
Gastric cancer cells%TNF-α%Lentiviral vector%Autocrine%inhibition function
目的：探讨TNF-α重组慢病毒对胃癌细胞自分泌杀伤作用。方法 TNF-α重组慢病毒感染SGC-7901胃癌细胞为实验组，并设立阴性对照组以及空白对照组，采用RT-PCR检测3组细胞中TNF-αmRNA的表达情况，ELISA检测3组细胞上清液中TNF-α蛋白含量，流式细胞术检测3组细胞凋亡情况。结果 PCR实验观察到TNF-αmRNA荧光条带24h后稳定表达。测得三组培养液中TNF-α含量，实验组[(2.6926±0.7563) ng/ml]明显高于阴性对照组[(0.9326±0.3091) ng/ml]和空白对照组[(0.8552±0.2768) ng/ml]，且差异均有统计学意义（P均<0.01），而阴性对照组和空白对照组之间差别无统计学意义（P>0.05）。在流式细胞实验中，实验组细胞凋亡率明显高于其他组，差异有统计学意义（P<0.05），而阴性对照组和空白对照组差别无统计学意义（P>0.05）。结论 TNF-α重组慢病毒可以感染SGC-7901胃癌细胞，并在胃癌细胞中稳定表达并自分泌TNF-α，诱导杀伤胃癌细胞。
目的：探討TNF-α重組慢病毒對胃癌細胞自分泌殺傷作用。方法 TNF-α重組慢病毒感染SGC-7901胃癌細胞為實驗組，併設立陰性對照組以及空白對照組，採用RT-PCR檢測3組細胞中TNF-αmRNA的錶達情況，ELISA檢測3組細胞上清液中TNF-α蛋白含量，流式細胞術檢測3組細胞凋亡情況。結果 PCR實驗觀察到TNF-αmRNA熒光條帶24h後穩定錶達。測得三組培養液中TNF-α含量，實驗組[(2.6926±0.7563) ng/ml]明顯高于陰性對照組[(0.9326±0.3091) ng/ml]和空白對照組[(0.8552±0.2768) ng/ml]，且差異均有統計學意義（P均<0.01），而陰性對照組和空白對照組之間差彆無統計學意義（P>0.05）。在流式細胞實驗中，實驗組細胞凋亡率明顯高于其他組，差異有統計學意義（P<0.05），而陰性對照組和空白對照組差彆無統計學意義（P>0.05）。結論 TNF-α重組慢病毒可以感染SGC-7901胃癌細胞，併在胃癌細胞中穩定錶達併自分泌TNF-α，誘導殺傷胃癌細胞。
목적：탐토TNF-α중조만병독대위암세포자분비살상작용。방법 TNF-α중조만병독감염SGC-7901위암세포위실험조，병설립음성대조조이급공백대조조，채용RT-PCR검측3조세포중TNF-αmRNA적표체정황，ELISA검측3조세포상청액중TNF-α단백함량，류식세포술검측3조세포조망정황。결과 PCR실험관찰도TNF-αmRNA형광조대24h후은정표체。측득삼조배양액중TNF-α함량，실험조[(2.6926±0.7563) ng/ml]명현고우음성대조조[(0.9326±0.3091) ng/ml]화공백대조조[(0.8552±0.2768) ng/ml]，차차이균유통계학의의（P균<0.01），이음성대조조화공백대조조지간차별무통계학의의（P>0.05）。재류식세포실험중，실험조세포조망솔명현고우기타조，차이유통계학의의（P<0.05），이음성대조조화공백대조조차별무통계학의의（P>0.05）。결론 TNF-α중조만병독가이감염SGC-7901위암세포，병재위암세포중은정표체병자분비TNF-α，유도살상위암세포。
Abstact:Objective To explore the expression of TNF-α gene by Lentiviral vector-mediated in SGC-7901 gastric cancer cells and the apoptosis induced by TNF-α that autocrined in gastric cancer cell.Methods The SGC-7901 gastric cancer cells were cultured. The gastric cancer cells were devided into three groups:1. the experimental group, the gastric cancer cells were infected with lentiviral vector-mediated TNF-αgene in cells, 2,the negative control group ,the gastric cancer cells infected with lentiviral vector cell group and 3,the control group ,SGC-7901 gastric cancer cells only. The expression of TNF-α was observed by PCR and Elisa method. The apoptosis of the gastric cancer cells were observed by flow cytometry.Results The expression of green fluorescent protein(GFP) was found in SGC-7901 gastric cancer cells infected with lentivirus 48h, and the expression efficiency was more than 90% when stable expression of the fluorescent trap on the TNF-α mRNA in experimental group after 24 hour. The TNF-α in the cultured liquid in the experimental group(2.6926±0.7563 ng/ml) was higher than that of in the negative control group (0.9326±0.3091 ng/ml) and in control group (0.8552±0.2768 ng/ml).T he differences were signiifcant (P<0.01).The differences was no signiifcant between the negative control group and the control group(P>0.05).Conclusion TNF-α genes mediated by lentivirus successfully infected gastric cancer cells and expressed TNF-α .The trans-TNF-α genes gastric cancer cells could autocrine TNF-α to induce apoptosis.
