LPLUNC1基因对鼻咽癌细胞HNE1生长增殖的抑制作用研究

LPLUNC1在正常的鼻咽组织及人胚鼻咽组织中高表达,而在71%的鼻咽癌中表达下调或缺失,是与鼻咽癌的发生发展密切相关的新基因.通过研究LPLUNC1基因对鼻咽癌细胞系HNE1的影响,进一步确定其与鼻咽癌发生发展的关系.将LPLUNC1基因全长cDNA克隆入pcDNA3.1( )真核表达载体中,通过脂质体介导稳定转染入LPLUNC1低表达鼻咽癌细胞系HNE1中,通过RT-PCR及RNA印迹筛选LPLUNC1高表达的细胞株,并利用细胞生长曲线、MTT、BrdU掺入、流式细胞仪检测、软琼脂集落形成实验及裸鼠成瘤等实验,研究了LPLUNC1对鼻咽癌细胞系HNE1细胞生长、增殖的影响.结果发现,稳定转染LPLUNC1的HNE1细胞的生长速度明显减慢,在MTT与BrdU掺入实验发现LPLUNC1可明显地抑制鼻咽癌细胞的增殖,并且通过流式细胞仪检测也发现,LPLUNC1基因可明显延缓HNE1细胞的细胞周期进程,使G0/G1期细胞增多而S期细胞相对减少.进一步通过软琼脂集落形成及裸鼠成瘤实验发现,LPLUNC1稳定转染后的HNE1细胞集落形成率与集落的大小均小于空白载体细胞,同时能明显地抑制HNE1细胞的体外成瘤.结果表明,LPLUNC1基因能明显抑制鼻咽癌细胞HNE1的生长增殖,是鼻咽癌发生发展中的重要候选抑瘤基因之一.

The Study of LPLUNC1 Gene Inhibit Human Nasopharyngeal Carcinoma Cell Line HNE1 Growth and Proliferation

LPLUNC1 is a newly cloned gene and was found down-expressed in 71% NPC biopsies but high-expressed in normal adult and fetal nasopharyngeal epithelial tissue, which indicates that it may play an important role in the tumorigenesis of nasopharyngeal carcinoma. The object of the research is to study the effect of LPLUNC1 on nasopharyngeal carcinoma cells. The full length of LPLUNC1 cDNA was cloned into the pcDNA3.1( ) vector, then the pcDNA3.1( )-LPLUNC1 plasmid was stably transfected into the NPC cell line HNE1 cells using lipofectamine. After being selected with G418 and detected with RT-PCR and Northern-blot, the stable transfectants of LPLUNC1 over-expression were constructed. Cell growth curve, MTT, BrdU labeling, flow cytometry, soft agar and nude mice were performed to study the effects of LPLUNC1 over-expression on HNE1 cells. The results showed that HNE1 cells with over-expression of LPLUNC1 grew slower significantly than that of the control. MTT and BrdU labeling assay showed that LPLUNC1 inhibited the HNE1 cell proliferation. And LPLUNC1 gene can delay G1-S phase cell cycle progression of HNE1 cells. Furthermore, LPLUNC1 was found that it decreased the HNE1 clonogenicity in soft agar and inhibited the tumor formation in nude mice obviously. In conclusion, LPLUNC1 can suppress the malignant phenotype of HNE1 and plays an important role in inhibiting the tumorigenesis and development of NPC.