Construction of recombinant lentivirus vector carrying lnc525893 and its effect on the proliferation of cervical cancer cells
ZHANG Mi1,2， DING Xiangya3， HU Minmin3， LI Wan3， LU Chun3， JIA Xuemei1,2
（1. Department of Gynecology and Obstetrics，the Fourth Clinical Medical College of Nanjing Medical University，Nanjing Jiangsu 210029； 2. Department of Gynecology and Obstetrics，Nanjing Maternity and Child Health Hospital Affiliated to Nanjing Medical University，Nanjing Jiangsu 210004; 3. Department of Microbiology，Nanjing Medical University，Nanjing Jiangsu 210029，China）
Abstract：[Abstract]Objective: To construct the recombinant lentivirus vector carrying long noncoding RNA 525893(lnc525893), and identify its effect on the proliferation of cervical cancer cells. Methods: The sequences of lnc525893 were cloned from human umbilical vein endothelial cells(HUVECs), and inserted into lentivirus vector pCDHCMVMCSEF1copGFP. Then recombinant plasmid pCDHlnc525893, package plasmid psPAX2 and envelope plasmid pMD2.G were cotransfected into the 293 T cells. Supernatant were harvested and filtered then the viral titer was determined by gradient dilution and counting cells which expressed the green fluorescent protein(GFP). Next, lentiviruslnc525893 and lentiviruspCDH were transduced into HeLa cells respectively, the expression of lnc525893 was detected by quantitative realtime PCR(qRTPCR). Cell counting kits assays were performed to identify its effect on the proliferation of HeLa cells. Results: The recombinant plasmid carrying lnc525893 was successfully constructed. The recombinant lentiviral titer was 1×107pfu/mL. The expression of lnc525893 in lnc525893HeLa cells was significantly higher than pCDHHeLa cells. In addition, overexpression of lnc525893 in HeLa cells inhibited the proliferation of HeLa cells. Conclusion: The recombinant lentivirus carrying lnc525893 was successfully constructed and lnc525893 could inhibit the proliferation of HeLa cells.
张咪1,2， 丁祥亚3， 胡敏敏3， 李婉3， 卢春3， 贾雪梅1,2. 长链非编码RNA525893重组慢病毒载体的构建及其对宫颈癌细胞增殖的影响[J]. 江苏大学学报:医学版, 2016, 26(02): 93-97.
ZHANG Mi1,2， DING Xiangya3， HU Min-min3， LI Wan3， LU Chun3， JIA Xue-mei1,2. Construction of recombinant lentivirus vector carrying lnc525893 and its effect on the proliferation of cervical cancer cells. Journal of Jiangsu University(Medicine Edition), 2016, 26(02): 93-97.
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2016, Vol. 26 
