结直肠癌中辅酶Q10的功能及分子机制探讨

Primary investigation on the function and molecular mechanism of coenzyme Q10 in colorectal cancer

摘要:

目的： 探索辅酶Q₁₀（coenzyme Q₁₀，CoQ₁₀）对结直肠癌细胞增殖和迁移能力的影响及潜在分子机制。方法： 使用CoQ₁₀分别处理SW480细胞和RKO细胞后，采用细胞计数和细胞划痕实验探究CoQ₁₀对结直肠癌细胞增殖和迁移能力的影响；采用流式细胞术探究CoQ₁₀对结直肠癌SW480细胞周期和细胞凋亡的影响；采用RNA-seq测序分析CoQ₁₀对结直肠癌RKO细胞系基因表达谱的影响，将差异表达基因进行GO和KEGG通路分析，最后采用定量RT-PCR验证RNA-seq结果。结果： ①CoQ₁₀对结直肠癌细胞系SW480的细胞增殖、细胞周期、细胞凋亡和细胞迁移均无明显影响，对结直肠癌细胞系RKO的细胞增殖也无明显影响，但可抑制RKO细胞迁移。②CoQ₁₀处理结直肠癌RKO细胞后的RNA-seq结果表明，差异表达基因明显富集于细胞黏附分子信号通路、细胞外基质受体信号通路和胆固醇信号通路等。定量RT-PCR验证结果表明，CoQ₁₀处理确实可导致胆固醇合成通路中的PCSK9、HMGCR、HMGCS1和DHCR7基因表达明显上调，提示CoQ₁₀可能通过调节这些基因的表达来抑制结直肠癌细胞RKO的迁移。结论： CoQ₁₀对结直肠癌细胞的增殖无明显影响，但可通过参与调节胆固醇代谢而抑制部分结直肠癌细胞的迁移能力。

Abstract:

Objective: To explore the potential molecular mechanism of coenzyme Q₁₀ (CoQ₁₀) on the proliferation and migration of colorectal cancer cells. Methods: SW480 cells and RKO cells were treated with CoQ₁₀ respectively, then cell count and cell scratch experiment were conducted to explore the effect of CoQ₁₀ on the proliferation and migration of colorectal cancer cells. SW480 cells were treated with CoQ₁₀ and then flow cytometry was performed to explore the effect of CoQ₁₀ on cell cycle and apoptosis of colorectal cancer SW480 cells. In order to explore the effect of CoQ₁₀ on gene expression profile of colorectal cancer RKO cell line, RKO cells treated with CoQ₁₀ were analyzed by RNA-seq sequencing, and the results were analyzed by GO and KEGG pathways. Finally, quanti-tative RT-PCR was used to verify the RNA-seq results. Results: ①CoQ₁₀ did not affect the proliferation, cell cycle, apoptosis and mi-gration of colorectal cancer cell SW480, nor did it affect the proliferation of colorectal cancer cell RKO, but it inhibited the migration of RKO. ②The RNA-seq results showed that differentially expressed genes were significantly enriched in cell adhesion molecule sig-naling pathway, extracellular matrix receptor signaling pathway and cholesterol signaling pathway. Quantitative RT-PCR results showed that CoQ₁₀ treatment did significantly up-regulate the expression of PCSK9, HMGCR, HMGCS1 and DHCR7 genes in cholesterol synthesis pathway. These results suggest that CoQ₁₀ may inhibit RKO migration in colorectal cancer cells by regulating the expression of these genes. Conclusion: CoQ₁₀ has no significant effect on the proliferation of colorectal cancer cells, but may inhibit the migra-tion ability of partial colorectal cancer cells by regulating cholesterol metabolism.