细胞周期蛋白D1反义寡核苷酸调控联合顺铂抑制口腔鳞癌耐药细胞的研究

Inhibition effect on Tca8113/CDDP by antisense oligonucleotides of cyclin D1 combined with cis diamminedichloroplatinum

目的通过体内外实验探讨细胞周期蛋白(cyclinD1)与舌鳞状细胞癌对顺铂(CDDP)耐药细胞系(Tca8113/CDDP)耐药的相关性。方法通过脂质体将cyclinD1正义、反义寡核苷酸序列分别导入Tca8113/CDDP细胞内。用甲基噻唑基四唑(MTT)法检测转染细胞系和耐药细胞系体外生长增殖情况及对顺铂的敏感性。将被转染的细胞及Tca8113/CDDP细胞接种于裸鼠皮下,建立移植瘤模型,共18只,每组6只。给予顺铂腹腔注射治疗,观察移植瘤体积变化,绘制生长曲线;称取瘤重,计算抑瘤率;光镜下观察移植瘤组织病理学变化。结果转导mRNA3′cyclinD1反义寡核苷酸的Tca8113/CDDP细胞在动物体内外生长明显受到顺铂的抑制;裸鼠皮下移植瘤抑瘤率达74%(P<0.05),移植瘤重量较对照组和转导正义寡核苷酸序列组差异均有统计学意义(P<0.05);光镜下可见反义寡核苷酸组移植瘤坏死明显。结论cyclinD1基因的反义调控联合顺铂能抑制Tca8113/CDDP细胞的生长能力及体内移植瘤的生长。研究结果提示:抑制cyclinD1可能增加口腔鳞状细胞癌对顺铂的敏感度。

Objective To clarify the relationship between cyclin D1 and cisplatin resistance of TcaS113/cis diamminedichloreplatinum（CDDP）in vitro and in vivo. Methods We applied the transfection method with plasmids pcDNA3.1-antisense-cyclin D1 by Lipofectamin 2000. TcaS113/CDDP cells were used as control. MTr assay was used to identify the proliferation and sensibility of those cells to cisplatin. Subsequently, 18 nude mice were subcutaneously injected by those cells and divided into 3 groups with 6 mice in each group. Every mouse was treated by cisplatin with 5 mg . kg^-1 . d^-1 for 5 days. The sizes of tumor were measured every other day and were described with the growth curves. After 20 days, tumors were anatomized and weighed. The tumor inhibition ratios were calculated and the HE slides were observed to determine the cell sensibility to cisplatin. Results The transfected cells with pcDNA3.1-antisense-cyclin Dlgrew more slowly than other cells and showed higher sensibility to cisplatin in vitro. The tumors developed by cells with pcDNA3.1-antisense-cyclin D1 were smaller than the The tumor inhibition ratio was 74% （P 〈 0. 05 ）. The necrosis area was larger in the tumors developed by the trausfected cells with pcDNA3.1- antisense-cyclin D1 than other groups. Conclusions Antisense oligonucleotides of cyclin D1 can improve the sensibility of TcaS113/CDDP cells to cisplatin and inhibit the growth of tumors.