KAI1基因抗胰腺癌细胞转移的研究

The studies on the role of KAI1 gene in anti-metastasis of primary pancreatic cancer cells

目的 将pCMV KAI1重组质粒转染入人高转移胰腺癌细胞株PANCⅠ和MiaPaCaⅡ中 ,观察转染前后KAI1基因对胰腺癌细胞增殖能力的影响 ,探讨KAI1基因抗胰腺癌转移的调控作用。方法 通过脂质体转染法将 pCMV KAI1重组质粒成功转染到人胰腺癌细胞株PANCⅠ和MiaPaCaⅡ中 ,采用MTT比色法、细胞集落形成和流式细胞术 (FCM )测定细胞增殖活力和生长周期的变化。结果 pCMV KAI1重组质粒转染PANCⅠ和MiaPaCaⅡ人胰腺癌细胞株后 ,细胞增殖能力较转染前分别降低 4 0 .5 9%和 6 5 .84 % (P <0 .0 5 )。转染KAI1基因的细胞集落形成率 (14 .0 0 % )较转染前 (4 9.5 6 % )明显减少 (P <0 .0 1)。转染后比转染前胰腺癌细胞数量在G0 /G1期明显增加 (PANCⅠ细胞由 4 0 .97%± 1.34%增加到 6 5 .36 %± 1.2 3% ,MiaPaCaⅡ细胞由 2 4 .10 %± 1.0 0 %增加到 4 3.18%± 1.0 6 % ) ,M/G2 期数量降低 (PANCⅠ从 2 1.4 6 %± 0 .96 %降至 13.35 %± 0 .2 2 % ,MiaPaCaⅡ由 2 4 .89%± 1.14 %降至 19.0 6 %± 0 .5 7% ) ,差异有显著性 (P <0 .0 5 )。结论 KAI1基因控制胰腺癌细胞转移可能是通过改变胰腺癌细胞的生长周期 ,抑制癌细胞的增殖及分裂功能来实现的。

Objective To evaluate the effect of recombinant expressing plasmid pCMV-KAI1 on the proliferative ability of PANC Ⅰ and MiaPaCa Ⅱ pancreatic cancer cells, and to observe the suppress metastatic mechanism of KAI1 gene in the malignancy.Methods The plasmid pCMV-KAI1 was transfected into the human pancreatic cancer cell lines PANC Ⅰ and MiaPaCa Ⅱ with liposome. The proliferative ability of these two pancreatic cancer cell lines were analyzed with MTT and colony-forming test, the cycle pattern was assayed by flow cytometry.Results After KAI1 expressing plasmid tranfected into PANC Ⅰ and MiaPaCa Ⅱ, the cell growth rates of the two cell lines were reduced by 40.59% and 65.84% respectively compared with the control cells (P<0.05). Colony formation in the transfected cells significantly decreased compared with the control cells (49.56% vs. 14.00%,P<0.01). Flow cytometry analysis revealed that the number of cells in G 0/G 1 phase increased markedly in KAI1-transfected PANC Ⅰ cells from (40.97±1.34)% to (65.36 ±1.23)% and in KAI1-transfected MiaPaCa Ⅱ cells from (24.10±1.00)% to (43.18±1.06)%. However, the number of cells in M/G 2 phase decreased, from (21.46±0.96)% to (13.35±0.22)% in KAI1-transfected PANC Ⅰ cells and from (24.89±1.14)% to (19.06±0.57)% in KAI1-transfected MiaPaCa Ⅱ cells (P<0.05).Conclusions The change of cycle pattern and the reduced proliferative ability of pancreatic cancer cells transfected with KAI1 gene may be the mechanism of KAI1 gene suppressing the metastasis of pancreatic cancer cell.