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. 2024 Aug 20;44(8):1497-1507.
doi: 10.12122/j.issn.1673-4254.2024.08.08.

[Biological role of SPAG5 in the malignant proliferation of gastric cancer cells]

[Article in Chinese]
Y Pang  1 , Y Liu  1 , S Chen  1 , J Zhang  1 , J Zeng  1 , Y Pan  2 , J An  1
Affiliations

[Biological role of SPAG5 in the malignant proliferation of gastric cancer cells]

[Article in Chinese]
Y Pang et al. Nan Fang Yi Ke Da Xue Xue Bao. .

Abstract
in English, Chinese

Objective: To analyze the expression of SPAG5 in gastric cancer tissues and its regulatory roles in gastric cancer cell growth.

Methods: TCGA analysis, immunohistochemistry, and immunofluorescence staining were used to analyze the expression patterns of SPAG5 and MKi67 in gastric cancer and adjacent tissues. In gastric cancer AGS and MGC803 cells, the effects of lentivirus-mediated SPAG5 knockdown on cell growth and apoptosis were evaluated using Celigo, MTT, clone formation assays and flow cytometry.

Results: Proteinatlas and TCGA database analysis suggested that SPAG5 was highly expressed in gastric cancer, and Kaplan-Meier analysis and GEPIA analysis showed high expressions of SPAG 5 in lung adenocarcinoma, breast cancer, hepatocellular carcinoma, pancreatic carcinoma, cervical cancer and bladder carcinoma. Immunohistochemistry revealed that SPAG5 was highly expressed in gastric cancer tissues (P < 0.001), and immunofluorescence colocalization analysis demonstrated a significant correlation between SPAG5 and MKI67 (R=0.393, P < 0.001). RT-qPCR and Western blotting showed that SPAG5 was highly expressed in MKN74, BGC823, MGC803, SGC7901 and AGS cells. In AGS and MGC803 cells, SPAG5 knockdown significantly inhibited proliferation and promoted apoptosis.

Conclusions: The expressions of SPAG5 and MKi67 are correlated in gastric cancer tissues, and SPAG5 knockdown inhibits the proliferation of gastric cancer cells. SPAG5 is associated with the prognosis of gastric cancer patients and may serve as a promising biomarker for gastric cancer.

目的: 在胃癌组织及癌旁组织中分析SPAG5的表达情况，通过干扰SPAG5分析其在胃癌细胞生长中的生物学作用。

方法: 结合TCGA分析，免疫组织化学、免疫荧光染色分析SPAG5及MKi67在胃癌及癌旁组织中的表达模式，利用胃癌细胞AGS和MGC803进行体外细胞生物学实验，分别设置空白对照组和干扰组，采用慢病毒干扰，其中空白对照转染shCtrl，干扰组转染shSPAG5。通过Celigo、MTT和克隆形成实验、凋亡检测分析敲减SPAG5基因后对胃癌细胞生长的影响。

结果: Proteinatlas数据库、TCGA数据库分析结果发现SPAG5在胃癌中高表达，KM-plot及GEPIA数据库分析SPAG5在肺腺癌、乳腺癌、肝癌、胰腺癌、宫颈癌、膀胱癌中高表达，免疫组化发现SPAG5在胃癌中高表达（P<0.001），免疫组化和免疫荧光共聚焦证明SPAG5与MKi67存在显著相关性（R=0.393，P<0.001）。Real time-PCR及Western blotting结果显示SPAG5在MKN74、BGC823、MGC803、SGC7901和AGS等细胞中表达水平较高（P<0.01）。敲减SPAG5后mRNA和蛋白的表达下降，Celigo、MTT和克隆形成实验结果显示敲减SPAG5抑制胃癌细胞增殖（P<0.01），流式凋亡分析发现敲减SPAG5促进胃癌细胞凋亡（P<0.001）。

结论: SPAG5和MKi67在胃癌组织中的表达具有相关性，干扰SPAG5基因可以抑制胃癌细胞的增殖。SPAG5与患者预后相关，可能成为胃癌的潜在标志物。

Keywords: MKi67; SPAG5; cell proliferation; gastric cancer.

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Figures

图1
图1
SPAG5表达模式的生物信息分析 Fig.1 Bioinformatic analysis of SPAG5 expression patterns. A:Proteinatlas database analysis showing SPAG5 distribution in both the cell nucleus and cytoplasm. B: TCGA database analysis showing significantly higher expression of SPAG5 in gastric cancer than in normal tissues (*P<0.05). C: Proteinatlas database analysis showing SPAG5 expressions in gastric, rectal adenocarcinoma, esophageal, colon adenocarcinoma, and pancreatic cancer.
图2
图2
KM-plot及GEPIA数据库分析SPAG5在多种实体肿瘤中的表达 Fig.2 Kaplan-Meier plots and GEPIA database analysis of SPAG5 expression in lung adenocarcinoma (A), breast cancer (B), hepatocellular carcinoma (C), pancreatic cancer (D), cervical cancer (E), and bladder cancer (F) and its association with patient survival (*P<0.05).
图 3
图 3
免疫组化和生存分析SPAG5 Fig.3 Immunohistochemistry for SPAG5 in gastric and adjacent tissues and survival analysis of SPAG5 expression. A: Immunohistochemistry for detecting expression of SPAG5 in gastric cancer and adjacent tissues (Original magnification: ×100). a: Normal gastric mucosal tissue Negative b: Normal gastric mucosal tissue Positive c: Gastric cancer tissue Negative d: Gastric cancer tissue Positive. B-D: Kaplan-Meier survival analysis of gastric cancer patients with high and low SPAG5 expression using Log-rank test and Breslow test.
图7
图7
干扰SPAG5后AGS和MGC803细胞的增殖受到显著抑制 Fig.7 Lentivirus-mediated SPAG5 interference inhibits proliferation of AGS and MGC803 cells. A, B: Celigo assay and MTT assays showing inhibition of proliferation of AGS and MGC-803 cells 3 days after lentivirus infection in the experimental group was significantly inhibited. C: Number of clones formed by AGS and MGC-803 cells with SPAG5 knockdown (×100). ***P<0.001.
图8
图8
干扰SPAG5促进AGS和MGC803细胞的细胞凋亡 Fig.8 SPAG5 interference promotes apoptosis in AGS and MGC803 cells. A, B: Scatter plot of flow cytometry after SPAG5 interference in AGS cells and MGC803 cells. C, D: Apoptosis rates in AGS cells and MGC803 cells 5 days after SPAG5 interference (***P<0.001).
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