Construction and cell function study of HHLA2-overexpressing gastric cancer cells

doi:10.3877/cma.j.issn.2095-2015.2023.06.002

Abstract

Abstract:

Objective

To construct gastric cancer cell lines overexpressing human endogenous retrovirus subfamily H long terminal repeat associating protein 2(HHLA2)by lentivirus vector and verify their biological functions.

Methods

The target gene and Lipo2000 reagent were used to construct recombinant plasmid, which was then applied to transfect 293T cell.We collect The cell supernatant was concentrated and the concentration was determined.The concentrated liquid was then transfected into gastric cancer cell lines BGC-823 and MGC80-3.The flow cytometry was used to sort and verify the GFP+ stable overexpressing cell lines.Cell function experiments(including migration experiment and proliferation experiment)were applied to explore the biological functions of overexpressing HHLA2 gastric cancer cell lines.

Results

The results of restriction enzyme digestion and sequencing showed that the recombinant lentivirus vector was successfully constructed.Flow sorting and verification showed that HHLA2 overexpressed gastric cancer stably transformed strains.Overexpression of HHLA2 could accelerate the proliferation of gastric cancer cell lines MGC80-3 and BGC-823, and their absorbance values at 24 h, 48 h and 72 h were higher than those of the control cell lines(P<0.05). Overexpression of HHLA2 could enhance the migration ability of gastric cancer cell line BGC-823, and its migration rate at 24 h and 48 h was higher than that of the control cell line(P<0.05). However, the migration distance of MGC80-3 cells did not increase significantly(P>0.05).

Conclusion

Overexpression of HHLA2 can promote the proliferation and migration ability of gastric cancer.

Key words: Gastric cancer, Human endogenous retrovirus subfamily H long terminal repeat associating protein 2, Lentiviral vector, Flow cytometry

Xiaojuan Han, Jiaqian Xu, Yulan Zhu, Ying Wang, Yuan Li, Jun Feng, Dong Shao. Construction and cell function study of HHLA2-overexpressing gastric cancer cells[J]. Chinese Journal of Digestion and Medical Imageology(Electronic Edition), 2023, 13(06): 373-377.

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URL: https://zhxhbyyxzz.cma-cmc.com.cn/EN/10.3877/cma.j.issn.2095-2015.2023.06.002

https://zhxhbyyxzz.cma-cmc.com.cn/EN/Y2023/V13/I06/373

Figures/Tables 6

References 16

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