annexinⅠ对胰腺癌细胞生物学行为影响的体内研究

In vivo effect of annexin Ⅰ down-regulation on the growth of human pancreatic cancer in nude mice

目的探讨annexinⅠ在胰腺癌发生过程中的作用。方法应用RNA干扰技术，敲低annexinⅠ基因在mRNA水平的表达，并经Western blot检测证实。分别接种人胰腺癌Suit-Ⅱ细胞及转染annexinⅠ—siRNA2、annexinⅠ—siRNA3、annexinⅠ—siRNAN的Suit-Ⅱ细胞，建立裸鼠胰腺癌移植瘤模型，观察胰腺癌细胞的成瘤能力、肿瘤生长速度的变化，测定肿瘤体积和瘤重。结果（1）Western blot检测结果显示，转染pSilencer—annexinⅠ—siRNA1的Suit-Ⅱ细胞annexinⅠ表达显著降低，转染pSilencer—annexinⅠ—siRNA2和pSilencer—annexinⅠ—siRNA3的Suit—Ⅱ细胞annexinⅠ的表达几乎被完全抑制。（2）接种转染annexinⅠ—siRNA2、annexinⅠ-siRNA3细胞组的肿瘤生长速度较接种亲本Suit-Ⅱ细胞组明显减慢，肿瘤生长被抑制，抑制率分别达76．6％和68．4％。接种肿瘤细胞后44d，接种转染annexinⅠ-siRNA2细胞组、接种转染annexinⅠ-siRNA3细胞组的瘤重分别为0．8987和0．8992g，显著低于接种亲本Suit-Ⅱ细胞组和接种转染annexinⅠ—siRNAN细胞组（分别为2．5866和2．4070g，P〈0．001）。结论annexinⅠ基因在胰腺癌发生过程中起到了促进胰腺癌细胞生长增殖、增强胰腺癌细胞成瘤能力的重要作用，可作为基因治疗的潜在靶点。

Objective To further explore the effect of annexin Ⅰ on the tumor growth of human pancreatic cancer in nude mice. Methods To knock down the expression of annexin Ⅰ in pancreatic carcinoma cells by RNAi. A nude mouse model of human pancreatic cancer was established by subcutaneous inoculation of human pancreatic cancer cell line Suit- Ⅱ cells. The effect of annexin Ⅰ on tumor growth was assessed by tumor growth curve and tumor weight records, and Westen blot and flow cytometry were used to examine the expression of annexin Ⅰ after annexin Ⅰ -knocking down. Results The results of Western blot revealed that the expression of annexin Ⅰ was significantly decreased in Suit-Ⅱ cells transfected with pSilencer-annexin Ⅰ -siRNA1, and almost completely inhibited in the cells transfected with pSilencer-annexin Ⅰ -siRNA2 and pSilencer-annexin Ⅰ -siRNA3. The growth of tumors transfected with annexin Ⅰ -siRNA2 and annexin Ⅰ -siRNA3 was inhibited by 76.6% and 68.4%, respectively, in comparison with that of tumor from the parent Suit-Ⅱ cells. At 44 days after tumor cell inoculation, the tumor weight was 0.8987 g （ transfected with annexin Ⅰ -siRNA2 ） and 0. 8992 g （ transfected with annexin Ⅰ -siRNA3 ）, significantly lower （P 〈0.001 ） than that of tumor from parent Suit- Ⅱ cells （2.5866 g） and transfected with annexin Ⅰ - siRNAN （2.4070 g）. Conclusion annexin Ⅰ promotes the growth and proliferation of pancreatic carcinoma cells in vivo and increases the ability of tumor formation in nude mice. The results of this study support that annexin Ⅰ may become a potential target in gene therapy for this disease.