Li Yu, Shang Peng, Qian Ai-Rong, Wang Li, Yang Yong, Chen Zhi-Nan
Department of Cell Biology, Fourth Military Medical University, Xi'an 710032, Shaanxi Province, China.
World J Gastroenterol. 2003 Oct;9(10):2174-7. doi: 10.3748/wjg.v9.i10.2174.
To study the inhibitory effects of antisense RNA of HAb18G/CD147 on invasion of hepatocellular carcinoma(HCC) cells in vitro.
Antisense RNA of HAb18G/CD147 vector PCI-asHAb18G was constructed by reversely inserting HAb18G/CD147 cDNA to eukaryotic expression vector PCI-neo. The HCC cell line HHCC was transfected by PCI-asHAb18G via cation liposome. Expression of HAb18G/CD147 of transfected cells selected by G418 (geneticin) was observed by immuno-histochemical SP staining and FACS (fluorescence activated cell sorting). Gelatin zymography was used to determine the effect of PCI-asHAb18G on reducing secretions of MMP-2 and MMP-9 of the transfected cells. Boyden chamber was employed to test the invasion of HCC cells in vitro.
The construction of antisense RNA vector PCI-asHAb18G was verified correct by partial nucleotide sequencing and restricted endonuclease digestion. The expression of HAb18G/CD147 in transfected HHCC was inhibited by PCI-asHAb18G. Secretions of MMP-2 and MMP-9 of transfected HHCC were reduced and the invasion of transfected HHCC was inhibited compared to HHCC, respectively.
Invasion of HCC cells can be inhibited by antisense RNA of HAb18G/CD147. HAb18G/CD147 may be used as a potential target of drugs for anti-invasion and metastasis of HCC.
研究HAb18G/CD147反义RNA对体外肝癌细胞侵袭的抑制作用。
将HAb18G/CD147 cDNA反向插入真核表达载体PCI-neo构建HAb18G/CD147反义RNA载体PCI-asHAb18G。通过阳离子脂质体将PCI-asHAb18G转染肝癌细胞系HHCC。用免疫组织化学SP染色和流式细胞术观察经G418筛选的转染细胞中HAb18G/CD147的表达。用明胶酶谱法检测PCI-asHAb18G对转染细胞MMP-2和MMP-9分泌减少的作用。采用Boyden小室检测体外肝癌细胞的侵袭能力。
经部分核苷酸测序和限制性内切酶酶切验证,反义RNA载体PCI-asHAb18G构建正确。PCI-asHAb18G抑制了转染的HHCC中HAb18G/CD147的表达。与HHCC相比,转染的HHCC中MMP-2和MMP-9的分泌减少,侵袭能力受到抑制。
HAb18G/CD147反义RNA可抑制肝癌细胞的侵袭。HAb18G/CD147可能作为肝癌抗侵袭和转移药物的潜在靶点。