Department of Pharmacy, People's Hospital of Rizhao, Rizhao, Shandong, PR China.
Department of Pharmacy, Rizhao Maternal and Child Health Care Hospital, Rizhao, Shandong, PR China.
Biochem Biophys Res Commun. 2019 Mar 19;510(4):508-514. doi: 10.1016/j.bbrc.2019.01.109. Epub 2019 Feb 5.
Due to the poor prognosis and high mortality (over 90%), Pancreas ductal adenocarcinoma (PDAC) is listed as the 7th leading cause of cancer-related death in the world, while gemcitabine sensitivity is key important in PDAC therapy. SNHG14 is thought to be an oncogene in cancer progression. However, the possible role of SNHG14 underlying the progress of the PDAC cell, specifically in gemcitabine resistance remains to be determined.
We analyzed the PDAC-related data collected from TCGA. PDAC cell line (SW1990) was used as in vitro model. RT-qPCR and western blot were used to detect the autophagy-related gene expression level. MTT and flow cytometry approaches were used to determine cell viability and apoptosis rate. The luciferase reporter assay was used to confirm the direct interaction between SNHG14 and miR-101. The wound healing assay and transwell assay were used to detect the migration and invasion abilities of PDAC cells.
The expression of SNHG14 was significantly higher in the PDAC tissues than in the normal tissues, while miR-101 was significantly downregulated in the PDAC tissues. Moreover, the correlation analysis showed that SNHG14 was negatively correlated with miR-101. The in vitro experiments furthermore confirmed their impacts on PDAC cells. Overexpression of SNHG14 and miR-101 inhibitor significantly enhanced cell proliferation, migration, and invasion rate of PDAC cell line. Moreover, SNHG14 knockdown and miR-101 mimics both led to attenuation of gemcitabine resistance-PDAC cell viability and promoted cell apoptosis rate, as well as the reduction of autophagy-related proteins (such as RAB5A and ATG4D). Overexpression of SNHG14 enhanced PDAC cell progression and inhibited cell apoptosis in gemcitabine treatment, as well as the increase of autophagy-related proteins, thus enhanced the chemoresistance of PDAC cells to gemcitabine.
Collectively, we first time revealed that SNHG14 could sponge miR-101 to enhance PDAC cell progression and find the specific axis of SNHG14/miR-101/autophagy underlying the chemoresistance in PDAC cells to gemcitabine, which could promote the progress of PDAC therapy.
由于预后不良和死亡率高（超过 90%），胰腺导管腺癌（PDAC）被列为全球第 7 大癌症相关死亡原因，而吉西他滨的敏感性是 PDAC 治疗的关键。SNHG14 被认为是癌症进展中的癌基因。然而，SNHG14 在 PDAC 细胞进展中的作用，特别是在吉西他滨耐药性方面，仍有待确定。
我们分析了来自 TCGA 的 PDAC 相关数据。SW1990 胰腺癌细胞系被用作体外模型。RT-qPCR 和 Western blot 用于检测自噬相关基因的表达水平。MTT 和流式细胞术用于检测细胞活力和细胞凋亡率。荧光素酶报告基因实验用于证实 SNHG14 和 miR-101 之间的直接相互作用。划痕愈合实验和 Transwell 实验用于检测 PDAC 细胞的迁移和侵袭能力。
PDAC 组织中 SNHG14 的表达明显高于正常组织，而 miR-101 在 PDAC 组织中明显下调。此外，相关性分析表明 SNHG14 与 miR-101 呈负相关。体外实验进一步证实了它们对 PDAC 细胞的影响。过表达 SNHG14 和 miR-101 抑制剂显著增强了 PDAC 细胞系的增殖、迁移和侵袭率。此外，SNHG14 敲低和 miR-101 模拟都导致吉西他滨耐药性-PDAC 细胞活力降低，并促进细胞凋亡率，以及自噬相关蛋白（如 RAB5A 和 ATG4D）减少。过表达 SNHG14 增强了吉西他滨治疗下的 PDAC 细胞进展并抑制了细胞凋亡，以及自噬相关蛋白的增加，从而增强了 PDAC 细胞对吉西他滨的化疗耐药性。
综上所述，我们首次揭示 SNHG14 可以通过海绵吸附 miR-101 来增强 PDAC 细胞的进展，并找到了 SNHG14/miR-101/自噬在 PDAC 细胞对吉西他滨耐药性中的具体作用轴，这可能促进了 PDAC 治疗的进展。
