Influence of Liposomal Transfection of Survivin Antisense Oligodeoxynucleotide on Human Pancreatic Cancer Cells_CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY

Authors：

LIU Haiwei ¹ , CHENG Le ¹ , WU Fei ¹ , LI Chengjie ¹ , CHEN Weiping ² , LIU Zhiming ¹

1.Department of Gastrointestinal Surgery, The First Affiliated Hospital of Guangxi Medical University, Nanning 530021, China;;
2.Department of Histology and Embryology, Guangxi Medical University, Nanning 530021, ChinaCorresponding Author: LIU Zhi-ming, E-mail: liuzhiminggy@163.com;

Corresponding author：

Keywords：

Pancreatic cancer; Survivin; Antisense; Oligonucleotide; Cell apoptosis

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Objective 　To investigate the inhibitory effect of survivin antisense oligonucleotides (ASODN) on proliferation of pancreatic cancer cells PANC-1.
Methods 　The ASODN and sense oligodeoxynucleotides (SODN) were complementary to survivin sequences. FAM-marked ASODN was transfected into PANC-1 cells mediated by positive ion liposome as ASODN group. Blank control group (normal cells), negative control group (normal medium)， and SODN group were established for comparison. The transfection efficiency was detected by flow cytometry (FCM) after transfection; MTT assay was used to detect cytotoxicity; Cell morphological changes were examined by transmission electron microscopy; The cell cycle and apoptotic rate were analyzed by FCM; Immunohistochemical staining techniques were used, and the expressions of survivin were observed under light microscopy, examined and analysed by computer image.
Results 　①The transfection efficiency was 31.9%, 37.4%, 41.4%, 52.6%, 　24.2%, 11.4%, 16.1%, and 15.5% when the transfecting concentration of ASODN was 50, 100, 150, 200, 250, 400, 600, and 800 nmol/L, respectively; The transfection efficiency was 12.0%, 50.8%, and 11.2% when the inoculated cells was 2×104/well, 2×105/well, and 2×106/well, respectively; The transfection efficiency was 　58.8%, 　34.0%, and 23.6% when 2 μl, 3 μl, and 4 μl liposome was used during transfection, respectively. ②Cell gap was oversize, morphous was round, adherent cells were less after transfection under fluorescence microscope. ③The inhibition rate in the ASODN group was higher than that in each control group (P lt;0.05) on 24, 36, 48 h after treating by survivin ASODN, which increased as time prolonged (P lt;0.05). ④The apoptosis showed a ladder-shaped line in the ASODN group. ⑤Apoptotic morphology was demonstrated in the ASODN group, such as apoptotic cells with nuclear chromatin highly concentrated, crescent nuclear staining aggregated by the side nuclear membrane, nucleolus disappeared by AO and EB stains. ⑥The apoptotic rate 〔（38.1±3.4）%〕 in the ASODN group was higher than that in the SODN group 〔（4.16±1.7）%〕, P lt;0.05. ⑦G2/M cell cycle arrested in the ASODN group. ⑧After transfection, the expression of survivin protein in the ASODN group was significantly lower than that of each control group (P lt;0.05).
Conclusions 　The optimal transfection conditions are as following: the cell count of 2×105/well, concentration of ASODN 200 nmol/L, and cationic liposome oligofectamine 2 μl, respectively. Survivin ASODN can inhibit the proliferation of pancreatic cancer cells and induce their apoptosis.

Citation： LIU Haiwei,CHENG Le,WU Fei,LI Chengjie,CHEN Weiping,LIU Zhiming. Influence of Liposomal Transfection of Survivin Antisense Oligodeoxynucleotide on Human Pancreatic Cancer Cells. CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY, 2010, 17(10): 1056-1061. doi: Copy

1.	Guo X, Cui Z. Current diagnosis and treatment of pancreatic cancer in China [J]. Pancreas, 2005; 31(1): 13-22.
2.	王丽，杨功焕，李辉，等. 1991-2000年中国胰腺癌病死率的变迁 [J]. 中华内科杂志， 2005； 44(7)： 509-513.
3.	Rodriguez JA, Li M, Yao Q, et al. Gene overexpression in pancreatic adenocarcinoma: diagnostic and therapeutic implications [J]. World J Surg, 2005; 29(3): 297-305.
4.	Devesa SS, Blot WJ, Stone BJ, et al. Recent cancer trends in the United States [J]. J Natl Cancer Inst, 1995; 87(3): 175-182.
5.	Ambrosini G, Adida C, Altieri DC. A novel anti-apoptosis gene, survivin, expressed in cancer and lymphoma [J]. Nat Med, 1997; 3(8): 917-921.
6.	Tamm I, Wang Y, Sausville E, et al. IAP-family protein survivin inhibits caspase activity and apoptosis induced by Fas (CD95), Bax, caspases, and anticancer drugs [J]. Cancer Res, 1998; 58(23): 5315-5320.
7.	Kawasaki H, Toyoda M, Shinohara H, et al. Expression of survivin correlates with apoptosis, proliferation, and angiogenesis during human colorectal tumorigenesis [J]. Cancer, 2001; 91(11): 2026-2032.
8.	Satoh K, Kaneko K, Hirota M, et al. Expression of survivin is correlated with cancer cell apoptosis and is involved in the development of human pancreatic duct cell tumors [J]. Cancer, 2001; 92(2): 271-278.
9.	谢贻祥, 孟刚. suvivin在乳腺癌中的表达及其意义 [J]. 中国普外基础与临床杂志, 2007; 14(3): 260-264.
10.	文亚渊, 刘宝华, 洪胜龙, 等. 凋亡相关基因survivin、caspase-3及cyclin-B1在胃癌发生、发展中的作用 [J]. 中国普外基础与临床杂志, 2006; 13(1): 34-37.
11.	Reed JC. The Survivin saga goes in vivo [J]. J Clin Invest, 2001; 108(7): 965-969.
12.	Li FZ, Ackermann EJ, Bennett CF, et al. Pleiotropic cell-division defects and apoptosis induced by interference with survivin function [J]. Nat Cell Biol, 1999; 1: 461-466.
13.	Carter BZ, Wang RY, Schober WD, et al. Targeting Survivin expression induces cell proliferation defect and subsequent cell death involving mitochondrial pathway in myeloid leukemic cells [J]. Cell Cycle, 2003; 2(5): 488-493.
14.	陈曦海，张岂凡，马荣. 反义survivin诱导胃癌SGC7901细胞凋亡及逆转耐药的研究 [J]. 中国普外基础与临床杂志, 2007; 14(1): 63-67.
15.	Xia C, Xu Z, Yuan X, et al. Induction of apoptosis in mesothelioma cells by antisurvivin oligonucleotides [J]. Mol Cancer Ther, 2002; 1(9): 687-694.

1. 　Guo X, Cui Z. Current diagnosis and treatment of pancreatic cancer in China [J]. Pancreas, 2005; 31(1): 13-22.
2. 　王丽，杨功焕，李辉，等. 1991-2000年中国胰腺癌病死率的变迁 [J]. 中华内科杂志， 2005； 44(7)： 509-513.
3. 　Rodriguez JA, Li M, Yao Q, et al. Gene overexpression in pancreatic adenocarcinoma: diagnostic and therapeutic implications [J]. World J Surg, 2005; 29(3): 297-305.
4. 　Devesa SS, Blot WJ, Stone BJ, et al. Recent cancer trends in the United States [J]. J Natl Cancer Inst, 1995; 87(3): 175-182.
5. 　Ambrosini G, Adida C, Altieri DC. A novel anti-apoptosis gene, survivin, expressed in cancer and lymphoma [J]. Nat Med, 1997; 3(8): 917-921.
6. 　Tamm I, Wang Y, Sausville E, et al. IAP-family protein survivin inhibits caspase activity and apoptosis induced by Fas (CD95), Bax, caspases, and anticancer drugs [J]. Cancer Res, 1998; 58(23): 5315-5320.
7. 　Kawasaki H, Toyoda M, Shinohara H, et al. Expression of survivin correlates with apoptosis, proliferation, and angiogenesis during human colorectal tumorigenesis [J]. Cancer, 2001; 91(11): 2026-2032.
8. 　Satoh K, Kaneko K, Hirota M, et al. Expression of survivin is correlated with cancer cell apoptosis and is involved in the development of human pancreatic duct cell tumors [J]. Cancer, 2001; 92(2): 271-278.
9. 　谢贻祥, 孟刚. suvivin在乳腺癌中的表达及其意义 [J]. 中国普外基础与临床杂志, 2007; 14(3): 260-264.
10. 　文亚渊, 刘宝华, 洪胜龙, 等. 凋亡相关基因survivin、caspase-3及cyclin-B1在胃癌发生、发展中的作用 [J]. 中国普外基础与临床杂志, 2006; 13(1): 34-37.
11. 　Reed JC. The Survivin saga goes in vivo [J]. J Clin Invest, 2001; 108(7): 965-969.
12. 　Li FZ, Ackermann EJ, Bennett CF, et al. Pleiotropic cell-division defects and apoptosis induced by interference with survivin function [J]. Nat Cell Biol, 1999; 1: 461-466.
13. 　Carter BZ, Wang RY, Schober WD, et al. Targeting Survivin expression induces cell proliferation defect and subsequent cell death involving mitochondrial pathway in myeloid leukemic cells [J]. Cell Cycle, 2003; 2(5): 488-493.
14. 　陈曦海，张岂凡，马荣. 反义survivin诱导胃癌SGC7901细胞凋亡及逆转耐药的研究 [J]. 中国普外基础与临床杂志, 2007; 14(1): 63-67.
15. 　Xia C, Xu Z, Yuan X, et al. Induction of apoptosis in mesothelioma cells by antisurvivin oligonucleotides [J]. Mol Cancer Ther, 2002; 1(9): 687-694.

CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY

Influence of Liposomal Transfection of Survivin Antisense Oligodeoxynucleotide on Human Pancreatic Cancer Cells

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