Influence of splicing factor MBNL3 on malignant biology behavior of pancreatic cancer cells

Abstract

Abstract: Objective To investigate MBNL3 expression and its effect on malignant biology behavior of pancreatic cancer cells. Methods Fifty paraffin samples of pancreas cancer excised during surgery in Peking Union Medical College Hospital from 2017 to 2020 were collected. Immunohistochemistry was conducted to explore expression of MBNL3. Pancreatic cancer cells were treated with MBNL3 knocked down or over-expressed by transfection. The proliferation and invasion or migration in different pancreas cells were examined respectively by real time cellular analysis and Transwell assay. The colony formation in different pancreas cells were examined by plate colony forming test. The MBNL3 expression in cell lines was analyzed by Western blot. Results Overall staining score of MBNL3 in pancreatic cancer tissues was 5.0±2.7, significantly higher than that of adjacent tissues(2.1±0.6)(P＜0.05). Cell proliferation experiment showed that 72 h after transfection, MBNL3 knockdown groups grew slower than control group in pancreatic cancer cells, whereas MBNL3 over-expressed groups grew faster than control group. Transwell assay showed that 10 h after transfection, invasion and migration of MBNL3 knockdown groups were weaker than control groups in pancreatic cancer cells (P＜0.05),meanwhile MBNL3 over-expressed groups were stronger than control groups in pancreaticcancer cells (P＜0.05). Colony forming experiments showed that colony formation number of MBNL3 knockdown groups were less than control groups in pancreatic cancer cells (P＜0.05), and the same of MBNL3 over-expressed groups were more than control group in pancreatic cancer cells(P＜0.05). Conclusions MBNL3 is highly expressed in pancreatic cancer tissue, which might take part in regulating the proliferation, invasion and migration of pancreatic cancer cells.

Key words: pancreatic cancer, splicing factor, invasion, migration

CLC Number:

R576

HAO Wen-zhen, CHEN Jie-min, FENG Yun-lu, WU Xi, WANG Qiang, WU Dong, YANG Ai-ming. Influence of splicing factor MBNL3 on malignant biology behavior of pancreatic cancer cells[J]. Basic & Clinical Medicine, 2021, 41(5): 688-693.

References

[1] Rahib L, Smith B, Rhonda A, et al. Projecting cancer incidence and deaths to 2030: the unexpected burden of thyroid, liver, and pancreas cancers in the United States[J]. Cancer Res, 2014, 74: 2913-2921.
[2] Zhang L, Sanagapalli S, Stoita A, et al. Challenges in diagnosis of pancreatic cancer[J]. World J Gastroenterol, 2018, 24: 2047-2060.doi:10.1038/s41598-020-58448-y.
[3] 由磊,陈革,赵玉沛. 胰腺癌干细胞研究进展[J]. 基础医学与临床,2009,29:885-887.
[4] Han H, Irimia M, Joel R,et al. MBNL proteins repress ES-cell-specific alternative splicing and reprogramming[J]. Nature, 2013, 498: 241-245.
[5] Yuan J, Liu X, Wang T, et al. The MBNL3 splicing factor promotes hepatocellular carcinoma by increasing PXN expression through the alternative splicing of lncRNA-PXN-AS1[J]. Nat Cell Biol, 2017, 19: 820-832.
[6] Grammatikakis I, Goo Y, Echeverria G, et al. Identification of MBNL1 and MBNL3 domains required for splicing activation and repression[J]. Nucleic Acids Res, 2011, 39: 2769-2780.
[7] Purcell J, Oddo J, Wang E, et al. Combinatorial mutagenesis of MBNL1 zinc fingers elucidates distinct classes of regulatory events[J]. Mol Cell Biol, 2012, 32: 4155-4167.
[8] Fardaei M, Rogers M, Thorpe H, et al. Three proteins, MBNL, MBLL and MBXL, co-localize in vivo with nuclear foci of expanded-repeat transcripts[J]. Hum Mol Genet, 2002, 11:805-814.
[9] Lee K, Cao Y, Witwicka H, et al. RNA-binding protein Muscleblind-like 3 (MBNL3) disrupts myocyte enhancer factor 2 (Mef2) {beta}-exon splicing[J]. J Biol Chem, 2010, 285: 33779-33787.
[10] Choi J, Dixon M, Dansithong W, et al. Muscleblind-like 3 deficit results in a spectrum of age-associated patho-logies observed in myotonic dystrophy[J]. Sci Rep, 2016, 6: 30999.doi:10.1038/srep30999.
[11] Tang Z, Li C, Kang B, et al. GEPIA: a web server for cancer and normal gene expression profiling and interac-tive analyses[J]. Nucleic Acids Res, 2017, 45:98-102.
[12] Luo G, Liu C, Guo M, et al. Potential biomarkers in Lewis negative patients with pancreatic cancer[J]. Ann Surg, 2017, 4: 800-805.

[1]	LIU Lei, LI Shi-bao, ZHANG Hao-liang. Lnc-SLC2A12-10:1 silencing inhibits the proliferation, migration and invasion of gastric cancer cell line AGS [J]. Basic & Clinical Medicine, 2022, 42(7): 1071-1076.
[2]	ZHONG Ming, YU Xiu-yi, GUO Wei-xi, FANG Zheng. Ophiopogonin B inhibits proliferation and migration of human esophageal squamous cell carcinoma cell line EC9706 [J]. Basic & Clinical Medicine, 2022, 42(7): 1053-1059.
[3]	BAI Bing, ZHANG Hai-fang, YANG Qing-liang, QIN Yue, ZHOU Chen-long, SONG Ge, WANG Ying. Down-regulation of lncRNA RHPN1-AS1 inhibits proliferation and migration of human bladder cancer cell line T24 [J]. Basic & Clinical Medicine, 2022, 42(6): 940-944.
[4]	DU Xue-mei, ZHANG Yun-mei, ZHOU Wen-ting, LYU Hong. miR-655-3p targeting KIF20A inhibits migration and invasion of lung adenocarcinoma cell line A549 [J]. Basic & Clinical Medicine, 2022, 42(4): 599-606.
[5]	CHEN Qiu-xia, YANG Li-xing, MA Rui, ZHAO Yong-jing, WANG Yu-cheng, JU Rui, GUO Lei. Carboxyamidotriazole-orotate inhibits proliferation and mitochondrial metabolism in human pancreatic cancer gemcitabine-resistant cell strain [J]. Basic & Clinical Medicine, 2022, 42(4): 570-576.
[6]	SUN Quan-peng, FAN Chao, ZHAN De-xi, FAN Yi-ming, SUN Wei-feng. Ropivacaine inhibits proliferation, migration and invasion of human liver cancer cell line Hep3B [J]. Basic & Clinical Medicine, 2022, 42(3): 448-453.
[7]	YANG Mang-zhuang, YANG Xu-dong, WANG Xiao-long. Circ-MALAT1 targeting miR-101 inhibits invasion and migration of bladder cancer cell line SW780 [J]. Basic & Clinical Medicine, 2022, 42(3): 472-478.
[8]	LIU Xin-bing, ZHANG Mei-hong. Gelsemine inhibits proliferation and invasion of rheumatoid arthritis synovial fibroblast through down-regulating circ_001680 [J]. Basic & Clinical Medicine, 2022, 42(3): 429-435.
[9]	CHEN Qian, LIU Xian, CUI Nan, ZHAI Yang. Knockout of EZH2 inhibits migration and invasion of human cervical cancer cell lines [J]. Basic & Clinical Medicine, 2022, 42(3): 395-400.
[10]	WANG Bu, YUAN Sheng-fang, ZHANG Chang-hong, YUAN Cheng, HUANG Pan-deng, ZHANG Zhi-hua, ZHAO Jian-qing. Silencing eEF2K combined with sodium tanshinone ⅡA sulfonate synergistically inhibit proliferation of human lung adenocarcinoma cell line A549 [J]. Basic & Clinical Medicine, 2022, 42(1): 106-113.
[11]	WANG Ling-xia, YAN Yu-lan, YUAN Hai-tao, CAI Ye-wei, LIU De-hui, GU Wen-lu, CAO Bi-yue. CircRNA_23113 inhibits the proliferation and migration of lung adenocarcinoma cells [J]. Basic & Clinical Medicine, 2022, 42(1): 82-88.
[12]	. Establishment of a serum-free culture method for human amnion mesenchymal stem cells [J]. Basic & Clinical Medicine, 2021, 41(8): 1181-1185.
[13]	. Zerumbone inhibits the proliferation, migration and invasion of human non-small cell lung cancer cell line A549 through up-regulating FBXW7 expression [J]. Basic & Clinical Medicine, 2021, 41(8): 1163-1168.
[14]	. Down-regulation of perilipin 2 inhibits proliferation of human gastric cancer cell line NCI-N87 [J]. Basic & Clinical Medicine, 2021, 41(7): 1013-1017.
[15]	. Macrophage J774A.1 exosome promotes the proliferation and migration of colorectal cancer cell lines MC38 and CT26 [J]. Basic & Clinical Medicine, 2021, 41(6): 799-804.