Establishment of pancreatic mucinous cystadenocarcinoma cell line MCC1 with stable overexpression of miR-224
10.3872/j.issn.1007-385x.2018.07.011
- VernacularTitle:稳定过表达miR-224的胰腺黏液性囊腺癌MCC1细胞株的建立
- Author:
PENG Xiaobo
1
;
GUO Chengtao
1
;
YING Mingzhen
1
;
LI Jie
1
;
SONG Lele
2
;
WU Yanjun
2
;
ZHAN Lixing
2
;
ZHAN Xianbao
1
Author Information
1. （1. Department of Oncology, Changhai Hospital, Second Military Medical University, Shanghai, 200433, China
2. Institute for Nutritional Science, Shanghai Institutes for Biological Sciences, ChineseAcademy of Sciences, Shanghai, 200031, China
- Publication Type:Journal Article
- Keywords:
miR-224;
lentiviral expression vector;
pancreatic mucinous cystadenocarcinoma
- From:
Chinese Journal of Cancer Biotherapy
2018;25(7):721-725
- CountryChina
- Language:Chinese
-
Abstract:
Objective: To construct a hsa-microRNA-224(miR-224) lentiviral expression vector and to establish pancreatic mucinous cystadenocarcinoma MCC1 cell line with stable miR-224 over-expression. Methods: Pri-miR-224 gene fragment was designed and amplified by quantitative real-time polymerase chain reaction (qRT-PCR), and then loaded into GV369 lentiviral vectors (GV369-miR224) by gene recombination technology. GV369-miR-224 lentivrial expression vectors were then identified by PCR and DNA sequencing. The GV369-miR-224 vector fluid was then used to infect pancreatic mucinous cystadenocarcinoma MCC1 cell line to establish the MCC1 cell line stably over-expressing miR-224. The transfection efficiency of GV369-NC and GV369-miR-224 was observed under fluorescence microscopy; and the expression levels of miR-224 in MCC1, GV369-miR-224-MCC1 and GV369-NC-MCC1 cell lines were detected by RT-PCR. Results: The GV369-miR-224 lentiviral vectors were successfully constructed. GV369-miR-224-MCC1 and GV369-NC-MCC1 cells all emit green fluorescence under the fluorescence microscope. The expression level of miR-224 in GV369miR-224-MCC1 cell group was significantly higher than that in negative control GV369-miR-224-MCC1 group and blank control MCC1 cell group (23.45±1.94， 1.46±0.1 and 2.11±0.38, P<0.01), however, there was no significant difference between the two control groups (P>0.05). Conclusion: A pancreatic mucinous cystadenocarcinoma MCC1 cell line with stable miR-224 over-expression was successfully established, and this will provide a new cell model for exploring the function and pathogenesis of miR-224 in pancreatic mucinous cystadenocarcinoma.
- Full text:20180711.pdf
