Effect of overexpression of KLF7 on proliferation and apoptosis of human non-small cell lung cancer cells

doi:10.3877/cma.j.issn.1674-6902.2025.01.001

Abstract

Abstract:

Objective

To investigate the regulation of KLF7 on the proliferation and apoptosis of lung cancer cells by constructing the KLF7 eukaryotic expression vector，thereby providing novel clues for the discovery of therapeutic targets for non-small cell lung cancer（NSCLC）.

Methods

The entire sequence of KLF7 was cloned through PCR，and the overexpression vector was constructed by means of enzyme digestion，ligation，and sequencing，with the expression efficiency verified.A594 cells were randomly divided into the experimental control group （control group） and the KLF7 overexpression group （KLF7 group）.Changes in the cell cycle，apoptosis，and reactive oxygen species of A594 cells were detected using flow cytometry，and the proliferation of A594 cells was examined by EdU.The expression levels of Proliferating Cell Nuclear Antigen（PCNA），G1/S-specific cyclin-D1 （CyclinD1），B-cell lymphoma-2 （Bcl-2，Bcl-2），Bcl-2-associated X protein （Bax） mRNA and protein were detected by real-time quantitative PCR and Western blotting.

Results

The total length of the CDS region of KLF7 was cloned to 693 bp and linked to the pCDNA3.1 vector to construct the KLF7 overexpression vector，which could significantly enhance the expression level of KLF7 in HEK293T and A594 cells （P＜0.01）.Compared with the control group，the percentage of cells in the G1 phase decreased，the percentage of cells in the G2 phase increased，the proliferation index （PI） significantly increased，and the percentage of EdU-positive cells significantly increased in the KLF7 overexpression group（P＜0.01）.Compared with the control group，the ROS level was inhibited and the apoptosis rate decreased in the KLF7 overexpression group （P＜0.01）.Compared with the control group，the relative expression levels of PCNA，CyclinD1，and Bcl-2 mRNA and protein increased in the KLF7 overexpression group，while the relative expression levels of Bax mRNA and protein decreased （P ＜0.01）.

Conclusion

In this study，the KLF7 overexpression vector was successfully constructed.In lung cancer cells，KLF7 overexpression can significantly promote cell proliferation and inhibit cell apoptosis，and participate in the occurrence of lung cancer，which may serve as a new biomarker of NSCLC.

Key words: Non-small cell lung cancer, Krüppel-like factor 7, Cells proliferation, Cells apoptosis, Lung cancer cells

Qiao Lv, Qian Min, Lu Jiang, Qian Chen, Jin Peng, Qian Dai. Effect of overexpression of KLF7 on proliferation and apoptosis of human non-small cell lung cancer cells[J]. Chinese Journal of Lung Diseases(Electronic Edition), 2025, 18(01): 1-7.

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References 53

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基因	引物序列（５′→３′）	片段长度（ｂｐ）	退火温度（℃）
ＫＬＦ７	Ｆ：ＡＧＣＧＡＣＡＴＧＡＧＣＡＡＧＧＡ	１５２	５８
	Ｒ：ＡＴＴＣＡＡＧＧＣＡＴＧＴＣＴＧＴＧＴＧ
β-ａｃｔｉｎ	Ｆ：ＧＡＴＡＴＴＧＣＴＧＣＧＣＴＣＧＴＧＧ	１８８	６０
	Ｒ：ＧＴＣＡＧＧＡＴＧＣＣＴＣＴＣＴＴＧＣＴ
ＰＣＮＡ	Ｆ：ＧＡＣＡＴＣＡＧＣＴＣＡＡＧＴＧＧＣＧ	１６２	５８
	Ｒ：ＧＣＡＧＣＧＧＴＡＡＧＴＧＴＣＧＡＡＧ
ＣｙｃｌｉｎＤ１	Ｆ：ＣＣＧＴＣＣＡＴＧＣＧＧＡＡＧＡＴＣ	１５６	６０
	Ｒ：ＣＡＧＧＡＡＧＣＧＧＴＣＣＡＧＧＴＡＧ
Ｂａｘ	Ｆ：ＧＡＣＡＣＡＧＡＣＴＣＴＣＣＣＣＧＡ	１７４	６１
	Ｒ：ＧＡＡＧＧＡＡＧＴＣＣＡＡＴＧＴＣＣＡ
Ｂｃｌ-２	Ｆ：ＧＴＧＧＡＴＧＡＣＣＧＡＧＴＡＣＣＴＧＡＡＣ	１９２	５９
	Ｒ：ＡＧＡＣＡＧＣＣＡＧＧＡＧＡＡＡＴＣＡＡＡＣ

基因	引物序列（５′→３′）	片段长度（ｂｐ）	退火温度（℃）
ＫＬＦ７	Ｆ：ＡＧＣＧＡＣＡＴＧＡＧＣＡＡＧＧＡ	１５２	５８
	Ｒ：ＡＴＴＣＡＡＧＧＣＡＴＧＴＣＴＧＴＧＴＧ
β-ａｃｔｉｎ	Ｆ：ＧＡＴＡＴＴＧＣＴＧＣＧＣＴＣＧＴＧＧ	１８８	６０
	Ｒ：ＧＴＣＡＧＧＡＴＧＣＣＴＣＴＣＴＴＧＣＴ
ＰＣＮＡ	Ｆ：ＧＡＣＡＴＣＡＧＣＴＣＡＡＧＴＧＧＣＧ	１６２	５８
	Ｒ：ＧＣＡＧＣＧＧＴＡＡＧＴＧＴＣＧＡＡＧ
ＣｙｃｌｉｎＤ１	Ｆ：ＣＣＧＴＣＣＡＴＧＣＧＧＡＡＧＡＴＣ	１５６	６０
	Ｒ：ＣＡＧＧＡＡＧＣＧＧＴＣＣＡＧＧＴＡＧ
Ｂａｘ	Ｆ：ＧＡＣＡＣＡＧＡＣＴＣＴＣＣＣＣＧＡ	１７４	６１
	Ｒ：ＧＡＡＧＧＡＡＧＴＣＣＡＡＴＧＴＣＣＡ
Ｂｃｌ-２	Ｆ：ＧＴＧＧＡＴＧＡＣＣＧＡＧＴＡＣＣＴＧＡＡＣ	１９２	５９
	Ｒ：ＡＧＡＣＡＧＣＣＡＧＧＡＧＡＡＡＴＣＡＡＡＣ

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