Effects of targeted silencing of PRL-3 gene on proliferation, migration, invasion and epithelial-mesenchymal transition of lung cancer cells

doi:10.3760/cma.j.cn371439-20200310-00003

Abstract

Abstract:

Objective To observe the effects of lentivirus-mediated shRNA silencing of PRL-3 gene on the proliferation, migration and invasion of lung cancer A549 cells and regulation of epithelial-mesenchymal transition (EMT) signaling pathway. Methods Lung cancer A549 cells were transfected with lentiviral interference vector carrying PRL-3 shRNA to build a stable PRL-3-silencing cell line. The cells were divided into blank control group, NC shRNA group (negative control group) and PRL-3 shRNA group (PRL-3 inhibiting RNAi lentivirus group). CCK-8 method, colony formation assay, Transwell and invasion chamber assay were performed to detect the proliferation, migration and invasion ability of A549 cells respectively. The expressions of E-cadherin and Snail mRNA were detected by real-time fluorescent quantitative PCR. Results The stable PRL-3-silencing cell line was successfully constructed. The knockdown efficiency of PRL-3 gene in the PRL-3 shRNA group reached 83.5%. CCK-8 method detected the proliferation ability of A549 cells, and the results showed the 24 h absorbance (A) values of A549 cells in the blank control group, NC shRNA group and PRL-3 shRNA group were 0.296±0.008, 0.342±0.007 and 0.292±0.004, with a statistically significant diffe-rence (F=106.300, P<0.001), and the PRL-3 shRNA group was significantly lower than the NC shRNA group (P<0.001); at 48, 72, 96 h after transfection, the cell proliferation abilities of the PRL-3 shRNA group were also significantly inhibited. Colony formation assay showed that the numbers of colony formation in the blank control group, NC shRNA group and PRL-3 shRNA group were 166.7± 6.7, 158.0±6.1 and 119.7±1.5 (F=67.290, P<0.001). The ability of colony formation of the PRL-3 shRNA group was significantly lower than that of the NC shRNA group (P<0.001). The numbers of migrated cells in the blank control group, NC shRNA group and PRL-3 shRNA group were 100.0±1.9, 98.8±1.9 and 44.6±7.6 (F=430.300, P<0.001). The migration ability of the PRL-3 shRNA group was significantly lower than that of the NC shRNA group (P<0.001). The numbers of invaded cells in the three groups were 117.7±4.1, 113.1±6.6 and 55.6±8.4 (F=247.200, P<0.001). The invasion ability of the PRL-3shRNA group was significantly lower than that of the NC shRNA group (P<0.001). Real-time fluorescent quantitative PCR detection results showed that after silencing the expression of PRL-3, the relative expression level of E-cadherin mRNA in A549 cells was significantly up-regulated, and the level of Snail mRNA was significantly down-regulated. Conclusion PRL-3 silencing can inhibit the proliferation, migration and invasion of A549 cells effectively. PRL-3 may affect the invasion of lung cancer cells through the EMT pathway.

Key words: Lung neoplasms, Phosphatase of regenerating liver-3, Invasion, Epithelial-mesenchymal transition

Wang Ningju, Chen Dongmei, Zhang Heng, Hu Ping, Wang Yan. Effects of targeted silencing of PRL-3 gene on proliferation, migration, invasion and epithelial-mesenchymal transition of lung cancer cells[J]. Journal of International Oncology, 2021, 48(1): 18-23.

Figures/Tables 6

References 14

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基因	引物序列(5'→3')
Snail	正向引物:CGGAAGCCTAACTACAGCGA
	反向引物:ACAGAGTCCCAGATGAGCATT
上皮钙黏素	正向引物:GGTTAAGCACAACAGCAACAG
	反向引物:GGCATCAGCATCAGTCACTT
GAPDH	正向引物:GTCTTCACCACCATGGAGAA
	反向引物:TAAGCAGTTGGTGGTGCAG

组别	24 h	48 h	72 h	96 h
空白对照组	0.296±0.008	0.391±0.006	0.949±0.030	1.555±0.025
NC shRNA组	0.342±0.007^a	0.395±0.011	0.957±0.030	1.540±0.020
PRL-3 shRNA组	0.292±0.004^b	0.347±0.003^b	0.660±0.025^b	1.121±0.054^b
F值	106.300	76.210	213.000	280.300
P值	<0.001	<0.001	<0.001	<0.001

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