Abstract：Objective：To observe the effects of baculovirus-mediated RNA small interference(siRNA) of DNA methyltransferase 1(DNMT1) on the apoptosis of pancreatic cancer cell line PaTu8988 cells. Methods: Human pancreatic cancer cell line PaTu8988 cells were cultured in 6-well plates. Pancreatic cancer cells were randomly divided into five groups: control group(C1 group), negative control group(C2 group), r-Bac-si-DNMT1-D1 group(D1 group), r-Bac-si-DNMT1-D2 group(D2 group), r-Bac-si-DNMT1-D3 group(D3 group). C1 group cells were cultured only with 10% fetal bovine serum(FBS) and DMEM. At 24 h after plating with FBS and DMEM, C2 group, D1 group, D2 group and D3 group were transfected with carrier control r-Bac-CMV-EGFP, and recombinant baculovirus r-Bac-si-DNMT1-D1, r-Bac-si-DNMT1-D2, r-Bac-si-DNMT1-D3, 150 μL each well. At 72 h after transfecting, flow cytometry was performed to analyze cell apoptosis. Western blotting was used to detect the expression of antiapoptosis protein Bcl-2 and proapoptosis protein Bax. Results: Flow cytometry analysis showed that cell apoptosis rate in D1 group, D2 group, and D3 group were significant higher than that in C1 group and C2 group(P<0.05). Compared with C1 group and C2 group, the protein expression of Bcl-2 in D1 group, D2 group and D3 group were deeply decreased(P<0.05), but the protein expression of Bax in D1 group, D2 group and D3 group were obviously higher(P<0.05). Conclusion: Baculovirus-mediated siRNA of DNMT1 could induce apoptosis of human pancreatic cancer cells by decreasing Bcl-2 expression and increasing Bax expression.
[1]Warsame R, Grothey A. Treatment options for advanced pancreatic cancer: a review [J]. Expert Rev Anticancer Ther, 2012, 12(10): 1327-1336.[2]Johnson AA, Akman K, Calimport SR, et al. The role of DNA methylation in aging, rejuvenation, and agerelated disease[J]. Rejuvenation Res, 2012, 15(5): 483-494.[3]张尤历，王晓燕， 徐岷，等. 杆状病毒介导的靶向DNA甲基转移酶1 siRNA表达载体的构建[J]. 江苏大学学报：医学版， 2011, 21(3): 237-240.[4]Zhang JJ, Zhu Y, Zhu Y, et al. Association of increased DNA methyltransferase expression with carcinogenesis and poor prognosis in pancreatic ductal adenocarcinoma [J]. Clin Transl Oncol, 2012, 14(2): 116-124.[5]Li A, Omura N, Hong SM, et al. Pancreatic cancer DNMT1 expression and sensitivity to DNMT1 inhibitors [J]. Cancer Boil Ther, 2010, 9(4): 321-329.[6]欧亮，张尤历，陈吉祥，等. 杆状病毒介导的DNMT1 siRNA对人胰腺癌裸鼠移植瘤的抑制作用[J]. 江苏大学学报:医学版， 2012, 22(3): 203-208.[7]Jia LT, Chen SY, Yang AG. Cancer gene therapy targeting cellular apoptosis machinery[J]. Cancer Treat Rev, 2012, 38(7): 868-876.[8]Nemec KN, Khaled AR. Therapeutic modulation of apoptosis: targeting the BCL2 family at the interface of the mitochondrial membrane[J]. Yonsei Med, 2008, 49(5): 689-697.[9]Wu M, Min C, Wang X, et al. Repression of BCL2 by the tumor suppressor activity of the lysyl oxidase propeptide inhibits transformed phenotype of lung and pancreatic cancer cells[J]. Cancer Res, 2007, 67(13): 6278-6285.[10]Bold RJ, Virudachalam S, McConkey DJ. BCL2 expression correlates with metastatic potential in pancreatic cancer cell lines[J]. Cancer, 2001, 92(5): 1122-1129.[11]Dumitru R, Gama V, Fagan BM, et al. Human embryonic stem cells have constitutively active Bax at the Golgi and are primed to undergo rapid apoptosis [J]. Mol Cell, 2012, 46(5): 573-583. [12]Sun Y, Zhao X, Yao Y, et al. Connexin 43 interacts with Bax to regulate apoptosis of pancreatic cancer through a gap junctionindependent pathway [J]. Int J Oncol, 2012, 41(3): 941-948.
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