MicroRNAs Affect Complement Regulator Expression and Mitochondrial Activity to Modulate Cell Resistance to Complement-Dependent Cytotoxicity

Abstract

MicroRNAs (miR) are small RNA molecules that shape the cell transcriptome and proteome through regulation of mRNA stability and translation. Here, we examined their function as determinants of cell resistance to complement-dependent cytotoxicity (CDC). To achieve this goal, we compared the expression of microRNAs between complement-resistant and -sensitive K562 leukemia, Raji lymphoma, and HCT-116 colorectal carcinoma cells. Global microRNA array analysis identified miR-150, miR-328, and miR-616 as regulators of CDC resistance. Inhibition of miR-150 reduced resistance, whereas inhibition of miR-328 or miR-616 enhanced cell resistance. Treatment of K562 cells with a sublytic dose of complement was shown to rapidly increase miR-150, miR-328, and miR-616 expression. Protein targets of these microRNAs were analyzed in K562 cells by mass spectrometry-based proteomics. Expression of the complement membrane regulatory proteins CD46 and CD59 was significantly enhanced after inhibition of miR-328 and miR-616. Enrichment of proteins of mitochondria, known target organelles in CDC, was observed after miR-150, miR-328, and miR-616 inhibition. In conclusion, miR-150, miR-328, and miR-616 regulate cell resistance to CDC by modifying the expression of the membrane complement regulators CD46 and CD59 and the response of the mitochondria to complement lytic attack. These microRNAs may be considered targets for intervention in complement-associated diseases and in anticancer, complementbased therapy.

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“…Among these, miR-328-5p was chosen for predicted candidate because not only it was the highest after transfecting with siRNA-lncRNA Glis2 (Fig. 6B) but also it was reported to be correlated with mitochondrial activity [24]. Data from qRT-PCR also revealed that the expression level of miR-328-5p is negatively correlated with lncRNA Glis2 (Fig.…”

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“…2 c-E). Moreover, CD46 and CD59 are the target molecules of miR-328, miR-616, and miR-20b/e, and at the same time, miR-155, miR-125b, and miR-9 target another regulatory factor of this system called complement factor H (CFH) all resulting in the excessive complement activation [214] , [218] , [219] , [220] ( Fig. 2 c-E).…”

“…2 c-E). Several miRNAs such as miR-150, miR-200b/c, and miR-217 underpin the more regulated activity of the complement system through the positive regulation of CD46 and CD55 molecules [214] , [221] ( Fig. 2 c-E).…”

“…Although incomplete MAC complexes do not cause cell lysis, they activate many cellular inflammatory responses ( e.g ., NF-κB pathway) and affect the production of pro-inflammatory cytokines, adhesion molecules, and miRNAs. Accordingly, the prompt increased expressions of miR-328 and miR-616 have been reported following the cell stimulation with sublytic doses of MAC that can have specific effects on the expression of their target molecules and consequently different cellular processes [214] ( Table 1 ). It has also been observed that C5a can change the expression of several miRNAs in the ECs.…”

“…Different complement components and regulators are among the target molecules of miRNAs. For instance, miR-145, miR-150, miR-218–2, and miR-92-d negatively regulate the C3 and miR-328 induces its deposition [213] , [214] , [215] , [216] . The miR-494-3p and miR-574 also target the C7 [217] ( Fig.…”