NKG5SV裸基因抑制裸鼠体内人肝癌细胞生长及转移

Injection of NKG5SV gene to inhibit growth and metastasis of hepatocellular carcinoma

目的 研究NKG5SV裸基因直接注射抗肝细胞肝癌生长及转移复发的作用。方法 以常规方法将NKG划SV装入逆转录病毒载体,扩增捉取NKG5裸基因,以LacZ为对照基因。将不同裸基因和生理盐水与LCI-D20裸鼠人肝癌同时皮下接种或待LCI-D20肿瘤成瘤后瘤内注射,观察对肿瘤成瘤和生长的影响。LCI-D20肿瘤裸鼠皮下接种后10d,分别瘤内注射不同裸基因,观察对肿瘤生长转移的影响。LC1-D20肿瘤肝内接种,接种后22d行肝癌切除术,术中经切缘或脾内注射不同裸基因,观察其对术后转移复发的影响。结累 不同裸基因与LCI-D20肝癌同时皮下接种对照组、LacZ组、NKG5SV组,肿瘤直径(cm)分别为1.76±0.11、1.51±0.34、0.33±0.04。裸基因肿瘤内注射对照组、LacZ细胞组、NKG5SV组,肿瘤直径(cm)分别为0.87±0.08、0.83±0.05、0.26±0.04。瘤重(g)分别为0.43±0.06、0.38±0.04、0.08±0.06。肝癌切除术中应用裸基因转移灶累及部位数、切缘复发瘤直径、肝内转移灶数目、转移灶累及肝叶数在对照组、LacZ组、NKG5SV组分别为,切缘注射肝外转移灶数目(个):4.25±1.48、4.25±1.04、0.63±0.51,切缘肿瘤直径(cm):1.51±0.27、1.35±0.17、0.81±0.17,肝内转移灶数目(个)2.50±1.41、2.38±1.06、1.25±0.71,转移灶累及肝叶数(个):2.13±0.99、2.

Objective To study the injection of NKG5SV gene to inhibit growth and metastasis of hepatocellular carcinoma (HCC). Methods NKG5SV gene was inserted into retroviral vector pLXSN by normal methods. LacZ gene was used as control. LCI-D20 tumor together with saline, pLXSN-LacZ DNA or pLXSN-NKG5SV was subcutaneously inoculated to the nude mice. Tumor formation rate and tumor size were noted 35 days after inoculation. LCI-D20 tumor was inoculated subcutaneously. Saline, pLXSN-LacZ DNA or pLXSN-NKGSSV was intratumorally injected respectively 10 days after inoculation. Tumor growth was observed 35 days after inoculation. Liver cancer was resected 22 days after intrahepatic inoculation. Saline, pLXSN-LacZ DNA or pLXSN-NKGSSV was respectively injected at incisal margin or intraspleen. Mice were killed 35 days after inoculation to observe tumor recurrence at incisal margin, intrahepatic metastasis and extrahepatic metastasis. Results Tumor formation rate and tumor diameter(cm) were 1.76 + 0.11, 1.51 + 0.34, 0.33 + 0.04 in the control group, LacZ group, NKG5SV group respectively when tumor and different cDNA were inoculated together. Tumor diameter (cm) and weight(g) were 0.87 +0.08, 0.83 +0.05, 0.26 +0.04; 0.43 + 0.06, 0.38 +0.04, 0.08 + 0.06 in the control group, LacZ group, NKG5SV group respectively when different cDNA were injected into the LCI-D20 tumor. Sites with extrahepatic metastasis nidi, incisal margin recurrence tumor size(cm), intrahepatic metastasis nidi, metastasis involved hepatic lobes in the control group, LacZ group, NKG5SV group were 4.25 + 1.48, 4.25 + 1.04, 0.63 + 0.51; 1.51 + 0.27, 1.35 + 0.17,0.81+0.17; 2.50+1.41, 2.38 + 1.06, 1.25 + 0.71; 2.13+0.99, 2.00+0.75, 1.38+0.74 respectively when NK cells were injected at incise margin. They were 4.38 +1.85, 4.25 + 1.48, 1.00 + 0.75; 1.13 + 0.23, 0.97 + 0.29, 0.76 + 0.16; 2.50 + 1.41, 2.05 + 1.12, 0; 2.13+0.83, 1.75+0.88, 0 respectively when NK cell were injected intrasplenicly. Conclusions NKG5SV gene can inhibit HCC growth and postoperative metastasis and recurrence.