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. 2023 Dec 13;23(1):507.
doi: 10.1186/s12890-023-02677-0.

Knockdown of MRPL35 promotes cell apoptosis and inhibits cell proliferation in non-small-cell lung cancer

Chengling Zhao #  1   2 , Lei Chen #  1   2 , Zhixin Jin  1   2 , Haitao Liu  1   2 , Chao Ma  1   2 , Hangtian Zhou  1   3 , Lingling Xu  1   2 , Sihui Zhou  1   3 , Yan Shi  1   3 , Wei Li  1   2 , Yuqing Chen  1   2 , Chengli Dou  4   5   6 , Xiaojing Wang  7   8   9
Affiliations

Knockdown of MRPL35 promotes cell apoptosis and inhibits cell proliferation in non-small-cell lung cancer

Chengling Zhao et al. BMC Pulm Med. .

Abstract

Background: Non-small cell lung cancer (NSCLC) is a major pathological type of lung cancer. However, its pathogenesis remains largely unclear. MRPL35 is a regulatory subunit of the mitoribosome, which can regulate the assembly of cytochrome c oxidases and plays an important role in the occurrence of NSCLC.

Methods: The expression of MRPL35 in NSCLC was detected by tissue microarray and immunohistochemistry. H1299 cells were infected with lentivirus to knockdown MRPL35, and the cells were subjected to crystal violet staining to assess the results of colony formation assays. A549 cells were infected by lentiviral particles-expressing shMRPL35 or shControl, and then subcutaneously injected into nude mice. Tumorigenesis in mice was detected by in vivo imaging. The potential pathway of MRPL35 in NSCLC was assessed by Western blotting.

Results: MRPL35 was over-expressed in NSCLC tissue compared to para-cancerous and normal tissues. Knockdown of MRPL35 suppressed cell proliferation and decreased NSCLC progression both in vitro and in vivo. The possible molecular mechanisms were also clarified, which indicated that MRPL35 could be involved in cell apoptosis and proliferation by modulating the expression levels of CDK1, BIRC5, CHEK1, STMN1 and MCM2. Knockdown of MRPL35 activated p53 signaling pathway and inhibited cell cycle regulation.

Conclusions: The oncogenic role of MRPL35 in NSCLC was potentially mediated through the cell cycle regulatory genes such as BIRC5, STMN1, CDK1, CHEK1 and MCM2, as well as activation of P53 signaling pathway.

Keywords: Apoptosis; MRPL35; NSCLC; Proliferation; p53.

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Conflict of interest statement

The authors declare no competing interests.

Figures

Fig. 1
Fig. 1
MRPL35 was over-expressed in NSCLC tissues. A IHC results of the expression level of MRPL35 in normal tissue, para-cancerous tissue, and NSCLC tissue. B Statistical analysis results of IHC staining intensity
Fig. 2
Fig. 2
MRPL35 knockdown promotes cell apoptosis and inhibits cell proliferation. A The mRNA level of MRPL35 in A549, NCI-H1299, 95-D, and NCI-H460 cells was detected by qPCR. B Colony formation assays. C Statistical analysis results of colony formation assays. D Cell proliferation was assessed by celigo cell count assays. E–F Statistical analysis results of celigo cell count assays. G Cell apoptosis in control and MRPL35-knockdown groups was detected by flow cytometry. H Statistical analysis results of flow cytometry
Fig. 3
Fig. 3
Knockdown of MRPL35 suppresses tumor growth in tumor-bearing mice. A In vivo imaging of tumor-bearing mice in control group and MRPL35-knockdown group. B Statistical analysis results of total radiant efficiency. C Tumor size was measured in NC and MRPL35-knockdown groups. D-E Measurement of the volume and weight of tumors
Fig. 4
Fig. 4
Transcriptome sequencing of NC group and MRPL35-knockdown group. A  Differentially expressed genes are illustrated using a volcano plot. B Heatmap showing the results of hierarchical clustering. C Canonical pathway enrichment analysis of differentially expressed genes. The red box marks the activation of the p53 signaling pathway and the inhibition of cyclins and cell cycle regulation
Fig. 5
Fig. 5
Interaction network of DEGs related to the cell proliferation and apoptosis. Red indicates the up-regulated genes after knockdown of MRPL35, green denotes the down-regulated genes, and different shapes represent different protein classifications. The red box marks the down-regulated genes related to cell proliferation and apoptosis after knockdown of MRPL35
Fig. 6
Fig. 6
Knockdown of MRPL35 results in the down-regulation of CHEK1, BIRC5, STMN1, MCM2 and CDK1. A-E The mRNA levels of CHEK1, BIRC5, STMN1, MCM2 and CDK1. F The protein levels of CHEK1, BIRC5, STMN1, MCM2 and CDK1. Full-length blots gels are presented in Supplementary Fig. 2
See this image and copyright information in PMC

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