Inhibitory effect and related mechanism of decitabine combined with gemcitabine on proliferation of NK/T cell lymphoma cells

Lanke Lin; Xiangqin Liu; Hui Yu; Huan Deng; Kun Peng; Jiang Chen; Chunle Zhang; Tao Jiang; Xiaoqi Liu

doi:10.3389/fphar.2023.1134895

Inhibitory effect and related mechanism of decitabine combined with gemcitabine on proliferation of NK/T cell lymphoma cells

Front Pharmacol. 2023 Mar 1:14:1134895. doi: 10.3389/fphar.2023.1134895. eCollection 2023.

Authors

Lanke Lin¹, Xiangqin Liu², Hui Yu³, Huan Deng¹, Kun Peng⁴, Jiang Chen⁵, Chunle Zhang⁶, Tao Jiang⁷, Xiaoqi Liu^{8

9}

Affiliations

¹ College of Medical Technology, Chengdu University of Traditional Chinese Medicine, Chengdu, China.
² Department of Laboratory Medicine and Sichuan Provincial Key Laboratory for Human Disease Gene Study, Sichuan Provincial People's Hospital, University of Electronic Science and Technology of China, Chengdu, China.
³ Department of Laboratory Medicine, The People's Hospital of Leshan, Leshan, China.
⁴ Health Management Center, Sichuan Provincial People's Hospital, University of Electronic Science and Technology of China, Chengdu, China.
⁵ The Department of Ophthalmology, Sichuan Provincial People's Hospital, University of Electronic Science and Technology of China, Chengdu, China.
⁶ Division of Nephrology, Kidney Research Institute, West China Hospital of Sichuan University, Chengdu, China.
⁷ Department of Hematology, Sichuan Academy of Medical Sciences & Sichuan Provincial People's Hospital, Chengdu, China.
⁸ Sichuan Provincial Key Laboratory for Human Disease Gene Study, Center for Medical Genetics, Sichuan Academy of Medical Sciences & Sichuan Provincial People's Hospital, University of Electronic Science and Technology of China, Chengdu, China.
⁹ Research Unit for Blindness Prevention of Chinese Academy of Medical Sciences (2019RU026), Sichuan Academy of Medical Sciences & Sichuan Provincial People's Hospital,, Chengdu, Sichuan, China.

Abstract

Background: EBV-associated lymphoma is a neoplasm with a poor prognosis, highly aggressive, and progressive rapidly. There is no standard clinical treatment protocol. Decitabine and gemcitabine are known to have anticancer properties against cells of various cancer, respectively. However, the effect of the combination medication on NK/T cell lymphoma cells and potential mechanisms have not been thoroughly investigated. Methods: Human NK/T cell lymphoma cells NK92MI were treated with decitabine and gemcitabine alone or in combination. Experiments, including the Cell Counting Kit-8 and flow cytometry, were performed to investigate how the combination of decitabine and gemcitabine affects the biological behavior of NK92MI cells in vitro. mRNA sequencing, RT-PCR, and western blotting were used to detect changes in the related signal pathway, mRNA, and protein expressions. Results: Decitabine and gemcitabine significantly inhibited the viability and proliferation of NK92MI cells in a dose-dependent manner. The combination index was less than 1 after treating with two drugs, which was a significant synergistic effect. The decitabine concentration with the best synergistic effect was 4.046 µM, and the gemcitabine concentration was 0.005 µM. Flow cytometry showed that combining two drugs could significantly promote apoptosis and arrest the cell cycle at the S phase. In the combined DAC and GEM group, caspase3 protein levels were higher than in either group alone or the control group. The transcriptome sequence, KEGG, and PPI analysis showed that the differential genes after combined treatment were mainly enriched in signal pathways related to cell proliferation, adhesion, and migration compared with using alone and control groups. Based on the sequencing results, we further investigated the role of DAC and GEM in ferroptosis-related signaling molecules using RT-PCR and Western blot techniques. RT-PCR and western blotting showed that the expression levels of HMOX1 and EBV cleavage gene BRLF1 were higher in the group with combined DAC and GEM than in the group alone and the control group, while the protein and mRNA expression levels of SLC7A11 were lower than the others. In addition, the GPX4 protein expression level in the combination group was lower than in the drug-alone and control groups. In addition, the combination treatment increased the ROS level of NK92MI cells. Conclusion: Our current findings suggested that decitabine had an inhibitory effect on the proliferation of NK92MI cells when co-treated with gemcitabine. This combination may increase the expression of ferroptosis-related signaling molecules, thus inhibiting the proliferation of NK92MI cells. It also promoted apoptosis in NK/T cell lymphoma. For patients with NK/T cell lymphoma, this novel combination may provide clinical benefits.

Keywords: NK/T cell lymphoma; apoptosis; decitabine; drug combination; ferroptosis; gemcitabine.

Grants and funding

This work was supported by the National Natural Science Foundation of China (82070930), the Sichuan Science and Technology Program (2022YFS0022, 2023YFS0308), the Chengdu Science & Technology Bureau (2021-YF05-02398-SN), Medico-Engineering Cooperation Funds from University of Electronic Science and Technology of China (ZYGX2021YGLH020) and the Foundation of Sichuan Medical Association (Q16069), Department of Science and Technology of Sichuan Provincial People's Hospital(2017LY15 and 2021LY07).