重楼皂苷VII通过SOHLH1诱导骨肉瘤铁死亡抑制肿瘤进展

doi:10.12122/j.issn.1673-4254.2025.12.20

南方医科大学学报 ›› 2025, Vol. 45 ›› Issue (12): 2726-2737.doi: 10.12122/j.issn.1673-4254.2025.12.20

• • 上一篇

重楼皂苷VII通过SOHLH1诱导骨肉瘤铁死亡抑制肿瘤进展

肖丹庭¹(), 唐海军¹, 杨明秀¹, 滕洪材¹, 梁积铭¹, 谢天裕², 冯文宇³, 刘尚玉¹, 戴薇¹, 李河柠⁴, 刘云¹()

^1.广西医科大学第一附属医院脊柱骨病外科，广西南宁 530021
^2.广西医科大学第一附属医院创伤骨科手外科，广西南宁 530021
^3.广西医科大学第二附属医院骨关节疾病外科，广西南宁 530021
^4.广西医科大学第一附属医院肿瘤科，广西南宁 530021

收稿日期:2025-08-10 出版日期:2025-12-20 发布日期:2025-12-22
通讯作者: 刘云 E-mail:szsyxdt@163.com;liuyun200450250@sina.com
作者简介:肖丹庭，在读硕士研究生，E-mail: szsyxdt@163.com
基金资助:
广西卫生健康委员会自筹研究项目(Z-A20230681)

Polyphyllin VII inhibits osteosarcoma xenograft growth in mice by inducing ferroptosis via upregulating SOHLH1

Danting XIAO¹(), Haijun TANG¹, Mingxiu YANG¹, Hongcai TENG¹, Jiming LIANG¹, Tianyu XIE², Wenyu FENG³, Shangyu LIU¹, Wei DAI¹, Hening LI⁴, Yun LIU¹()

^1.Department of Spinal Bone Disease, First Affiliated Hospital of Guangxi Medical University, Nanning 530021, China
^2.Department of Traumatic Orthopedics and Hand Surgery, First Affiliated Hospital of Guangxi Medical University, Nanning 530021, China
^3.Department of Bone and Joint Diseases Surgery, Second Affiliated Hospital of Guangxi Medical University, Nanning 530007, China
^4.Department of Oncology, First Affiliated Hospital of Guangxi Medical University, Nanning 530021, China

Received:2025-08-10 Online:2025-12-20 Published:2025-12-22
Contact: Yun LIU E-mail:szsyxdt@163.com;liuyun200450250@sina.com

摘要/Abstract

摘要：

目的探讨重楼皂苷VII（PP7）的抗骨肉瘤疗效及其分子机制。方法采用超高效液相色谱串联质谱分析鉴定重楼的主要成分。建立骨肉瘤患者来源异种移植瘤（PDX）模型，将构建的6只PDX模型的裸鼠随机分为实验组和对照组，3只/组。实验组接受2 mg/kg PP7灌胃（2 d/次，共用药28 d），对照组给予等量生理盐水，测量两组肿瘤的体积和质量。在143B和HOS细胞中，采用CCK-8法检测不同浓度PP7（0、1.25、2.5、5、10 μmol/L）对肿瘤细胞增殖的影响并确定起效浓度。通过Transwell实验分析PP7对细胞迁移和侵袭的作用。结合单细胞转录组测序（scRNA-seq）、批量转录组测序（bulk RNA-seq）和分子对接预测PP7的作用靶点，采用Western blotting验证PP7对该靶点的调控作用。构建沉默SOHLH1的骨肉瘤细胞，设置对照组、PP7干预组（5 μmol/L）、沉默SOHLH1组及PP7+沉默SOHLH1组，分别检测各组细胞迁移、侵袭能力以及铁死亡相关指标ROS和LPO的水平。结果质谱分析发现PP7为重楼主要活性成分之一。体内实验中，PP7处理组肿瘤体积与质量均显著低于对照组（P<0.05）。体外实验显示，PP7以浓度依赖方式抑制骨肉瘤细胞增值，并在5 μmol/L浓度下抑制细胞增殖（P<0.05），且该浓度抑制细胞迁移与侵袭（P<0.05）。多组学分析确定SOHLH1为PP7潜在作用靶点。Western blotting证实PP7可上调SOHLH1 mRNA及蛋白表达（P<0.05）。SOHLH1沉默削弱了PP7对细胞迁移、侵袭的抑制作用，并减弱了PP7诱导的铁死亡（P<0.05）。结论 PP7通过上调SOHLH1表达，诱导骨肉瘤细胞铁死亡，进而抑制其恶性进展。

关键词: 骨肉瘤, 重楼皂苷VII, 铁死亡, 单细胞转录组测序, PDX模型

Abstract:

Objective To investigate the inhibitory effect of polyphyllin VII (PP7) on osteosarcoma xenograft growth in mice and explore the underlying molecular mechanism. Methods Ultra‑performance liquid chromatography‑tandem mass spectrometry was used to analyze the main active components of Paris polyphylla. Six nude mice bearing patient‑derived xenograft (PDX) were randomized into two groups for treatment with 2 mg/kg PP7 gavage or saline every other day for 28 days, and the changes in tumor volume and mass were measured. In cultured 143B and HOS cells, the effect of PP7 treatment (0, 1.25, 2.5, 5, and 10 μmol/L) on cell proliferation was assessed with CCK‑8 assay, and Transwell assays were employed to examine the changes in cell migration and invasion. The target of PP7 was predicted by integrated analyses with single‑cell RNA sequencing (scRNA‑seq), bulk RNA sequencing (bulk RNA‑seq) and molecular docking and verified using Western blotting. In osteosarcoma cells transfected with SOHLH1 siRNAs or a negative control sequence, the effects of PP7 treatment (5 μmol/L) on cell migration, invasion, ferroptosis, reactive oxygen species (ROS) production and lipid peroxidation (LPO) were analyzed. Results PP7 was identified as one of the major active constituents of Paris polyphylla. In the tumor-bearing mice, PP7 treatment significantly lower the tumor volume and mass. In 143B and HOS cells, PP7 concentration‑dependently inhibited cell proliferation, and at 5 μmol/L, PP7 significantly inhibited cell proliferation, migration and invasion. Multi‑omics analysis identified SOHLH1 as a potential target of PP7, and Western blotting confirmed that PP7 upregulated SOHLH1 expressions at both the mRNA and protein levels. SOHLH1 silencing obviously attenuated the inhibitory effects of PP7 on cell migration and invasion and reduced PP7‑induced ferroptosis. Conclusion PP7 suppresses osteosarcoma xenograft growth in mice by inducing ferroptosis via upregulating SOHLH1 expression.

Key words: osteosarcoma, polyphyllin VII, ferroptosis, single-cell RNA sequencing, PDX model

肖丹庭, 唐海军, 杨明秀, 滕洪材, 梁积铭, 谢天裕, 冯文宇, 刘尚玉, 戴薇, 李河柠, 刘云. 重楼皂苷VII通过SOHLH1诱导骨肉瘤铁死亡抑制肿瘤进展[J]. 南方医科大学学报, 2025, 45(12): 2726-2737.

Danting XIAO, Haijun TANG, Mingxiu YANG, Hongcai TENG, Jiming LIANG, Tianyu XIE, Wenyu FENG, Shangyu LIU, Wei DAI, Hening LI, Yun LIU. Polyphyllin VII inhibits osteosarcoma xenograft growth in mice by inducing ferroptosis via upregulating SOHLH1[J]. Journal of Southern Medical University, 2025, 45(12): 2726-2737.

图/表 9

参考文献 36

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No.	Retention time (min)	Accurate molecular weight	Ionic form	ppm	Compound name	Molecular formula	First and second level fragments
1	1.798	402.4792	M+K	-3.17	Acetyl tributyl citrate	C20H34O8	441.5761, 199.0983, 215.0932, 229.142, 257.1379, 259.1531, 267.123, 287.1467, 299.1497, 303.18014
2	1.982	610.5175	M+H	-1.96	Rutin	C27H30O16	611.5242, 123.0442, 245.0442, 593.146,
3	4.618	274.3978	M+H	-2.54	Nandrolone	C18H26O2	275.4051, 83.0852, 131.0843, 153.1279, 177.1264, 275.2006
4	14.503	855.017	M+H	2.57	Polyphyllin I	C44H70O16	856.0271, 428.5142,
5	21.452	592.7606	M+H	1.35	Agavoside A	C33H52O9	593.7685, 123.1132, 291.1961, 475.3434, 519.3343,
6	21.687	903.0583	M+H	7.96	Trigofoenoside A	C45H74O18	904.059, 577.3734, 683.4028, 753.4484, 829.4611
7	25.211	293.3663	M+Na	4.43	Anastrozole	C17H19N5	316.3553, 55.0295, 240.1509, 173.1073, 187.1236, 225.1122
8	25.474	738.9018	M+H	-1.89	Polyphyllin VI	C39H62O13	739.9083, 433.2632, 578.383, 397.3106
9	27.384	430.62	M+H	3.94	Pennogenin	C27H42O4	431.6296
10	27.499	414.6206	M+H	0.96	Diosgenin	C27H42O3	415.6289, 81.0693, 105.0697
11	28.371	480.6341	M+H	9.34	Crustecdysone	C27H44O7	481.6467, 59.0493, 127.1128, 279.1596, 305.1737, 307.1907,
12	28.877	191.1867	M+H	5.72	Carbendazim	C9H9N3O2	192.1957
13	29.455	1015.1848	M+H	-2.85	Polyphyllin B	C51H82O20	1016.1898, 293.1246, 723.4232
14	29.455	1015.1848	M+H	-2.85	Polyphyllin E	C51H82O20	1016.1898
15	29.455	1015.1848	M+H	5.51	Polyphyllin F	C51H82O20	1016.1983
16	30.536	869.0436	M+H	1.72	Dioscin	C45H72O16	870.0534, 89.0600, 121.1013, 127.0393, 147.0616, 249.1868, 291.1082, 457.3314
17	32.939	696.8651	M+H	9.03	Momordicoside E	C37H60O12	697.8793, 489.3545
18	33.402	404.4105	M+H	0.34	Kaempferol 7-O-Neohesperidoside	C27H30O15	595.5262, 223.0970, 373.1257
19	37.217	323.5133	M+H	-0.31	Linoleoyl Ethanolamide	C20H37NO2	324.5211, 83.0849
20	37.434	278.3435	M+K	-1.89	Dibutyl Phthalate	C16H22O4	317.4412
21	37.434	317.5072	M+H	0.63	Phytosphingosine	C18H39NO3	318.5153, 65.038, 69.0704
22	38.378	1049.1990	M+H	-4.00	Polyphyllin VII	C51H84O22	1050.2032, 721.4158, 575.3579,
23	51.839	196.2429	M+H	3.55	Loliolide	C11H16O3	197.2516, 67.0547, 111.0446

No.	Retention time (min)	Accurate molecular weight	Ionic form	ppm	Compound name	Molecular formula	First and second level fragments
1	1.798	402.4792	M+K	-3.17	Acetyl tributyl citrate	C20H34O8	441.5761, 199.0983, 215.0932, 229.142, 257.1379, 259.1531, 267.123, 287.1467, 299.1497, 303.18014
2	1.982	610.5175	M+H	-1.96	Rutin	C27H30O16	611.5242, 123.0442, 245.0442, 593.146,
3	4.618	274.3978	M+H	-2.54	Nandrolone	C18H26O2	275.4051, 83.0852, 131.0843, 153.1279, 177.1264, 275.2006
4	14.503	855.017	M+H	2.57	Polyphyllin I	C44H70O16	856.0271, 428.5142,
5	21.452	592.7606	M+H	1.35	Agavoside A	C33H52O9	593.7685, 123.1132, 291.1961, 475.3434, 519.3343,
6	21.687	903.0583	M+H	7.96	Trigofoenoside A	C45H74O18	904.059, 577.3734, 683.4028, 753.4484, 829.4611
7	25.211	293.3663	M+Na	4.43	Anastrozole	C17H19N5	316.3553, 55.0295, 240.1509, 173.1073, 187.1236, 225.1122
8	25.474	738.9018	M+H	-1.89	Polyphyllin VI	C39H62O13	739.9083, 433.2632, 578.383, 397.3106
9	27.384	430.62	M+H	3.94	Pennogenin	C27H42O4	431.6296
10	27.499	414.6206	M+H	0.96	Diosgenin	C27H42O3	415.6289, 81.0693, 105.0697
11	28.371	480.6341	M+H	9.34	Crustecdysone	C27H44O7	481.6467, 59.0493, 127.1128, 279.1596, 305.1737, 307.1907,
12	28.877	191.1867	M+H	5.72	Carbendazim	C9H9N3O2	192.1957
13	29.455	1015.1848	M+H	-2.85	Polyphyllin B	C51H82O20	1016.1898, 293.1246, 723.4232
14	29.455	1015.1848	M+H	-2.85	Polyphyllin E	C51H82O20	1016.1898
15	29.455	1015.1848	M+H	5.51	Polyphyllin F	C51H82O20	1016.1983
16	30.536	869.0436	M+H	1.72	Dioscin	C45H72O16	870.0534, 89.0600, 121.1013, 127.0393, 147.0616, 249.1868, 291.1082, 457.3314
17	32.939	696.8651	M+H	9.03	Momordicoside E	C37H60O12	697.8793, 489.3545
18	33.402	404.4105	M+H	0.34	Kaempferol 7-O-Neohesperidoside	C27H30O15	595.5262, 223.0970, 373.1257
19	37.217	323.5133	M+H	-0.31	Linoleoyl Ethanolamide	C20H37NO2	324.5211, 83.0849
20	37.434	278.3435	M+K	-1.89	Dibutyl Phthalate	C16H22O4	317.4412
21	37.434	317.5072	M+H	0.63	Phytosphingosine	C18H39NO3	318.5153, 65.038, 69.0704
22	38.378	1049.1990	M+H	-4.00	Polyphyllin VII	C51H84O22	1050.2032, 721.4158, 575.3579,
23	51.839	196.2429	M+H	3.55	Loliolide	C11H16O3	197.2516, 67.0547, 111.0446

No.	Retention time (min)	Accurate molecular weight	Ionic form	ppm	Compound name	Molecular formula	First and second level fragments
1	6.609	402.4792	M-H	8.97	Acetyl Tributyl Citrate	C20H34O8	401.4748, 245.103, 255.1215, 289.0943
2	7.238	430.62	M-H	-1.40	Pennogenin	C27H42O4	429.6114
3	10.321	610.5175	M+Cl	8.36	Rutin	C27H30O16	645.9759, 163.0600, 193.0131
4	11.516	414.6206	M+Cl	3.55	Diosgenin	C27H42O3	450.0752
5	14.868	738.9018	M+Cl	4.52	Polyphyllin VI	C39H62O13	774.3583
6	21.72	592.7606	M-H	7.60	Agavoside A	C33H52O9	591.7571, 73.0284, 315.2324, 355.2267, 359.2231, 413.3022
7	26.119	191.1867	M-H	1.58	Carbendazim	C9H9N3O2	190.1790, 104.0244, 158.0367
8	29.339	594.5181	M-H	0.74	Kaempferol 7-O-Neohesperidoside	C27H30O15	593.5102, 111.0446, 251.0915, 265.0710, 375.1415
9	29.917	696.8651	M+Cl	2.32	Momordicoside E	C37H60O12	732.3134, 121.0515, 341.2867, 505.3542, 605.381
10	34.047	317.5072	M-H	-8.21	Phytosphingosine	C18H39NO3	316.4966, 352.9604
11	34.395	480.6341	M+Cl	0	Crustecdysone	C27H44O7	516.0871, 75.0456, 99.0812, 115.0761, 141.0934, 145.0868, 291.1613, 301.1825, 445.2586
12	36.535	903.0583	M+Cl	0.53	Trigofoenoside A	C45H74O18	938.5118, 547.3687, 593.3664, 709.4143
13	40.653	196.2429	M-H	6.15	Loliolide	C11H16O3	195.2362, 179.0700, 195.1020
14	41.316	278.3435	M+Cl	-6.05	Dibutyl Phthalate	C16H22O4	313.7946, 55.0548, 147.0078, 167.034, 183.1388
15	42.043	869.0436	M-H	-3.34	Dioscin	C45H72O16	868.0327, 101.0238, 557.3476, 575.3569
16	45.297	274.3978	M-H	5.49	Nandrolone	C18H26O2	273.3914, 147.0822, 173.0955, 239.1447

No.	Retention time (min)	Accurate molecular weight	Ionic form	ppm	Compound name	Molecular formula	First and second level fragments
1	6.609	402.4792	M-H	8.97	Acetyl Tributyl Citrate	C20H34O8	401.4748, 245.103, 255.1215, 289.0943
2	7.238	430.62	M-H	-1.40	Pennogenin	C27H42O4	429.6114
3	10.321	610.5175	M+Cl	8.36	Rutin	C27H30O16	645.9759, 163.0600, 193.0131
4	11.516	414.6206	M+Cl	3.55	Diosgenin	C27H42O3	450.0752
5	14.868	738.9018	M+Cl	4.52	Polyphyllin VI	C39H62O13	774.3583
6	21.72	592.7606	M-H	7.60	Agavoside A	C33H52O9	591.7571, 73.0284, 315.2324, 355.2267, 359.2231, 413.3022
7	26.119	191.1867	M-H	1.58	Carbendazim	C9H9N3O2	190.1790, 104.0244, 158.0367
8	29.339	594.5181	M-H	0.74	Kaempferol 7-O-Neohesperidoside	C27H30O15	593.5102, 111.0446, 251.0915, 265.0710, 375.1415
9	29.917	696.8651	M+Cl	2.32	Momordicoside E	C37H60O12	732.3134, 121.0515, 341.2867, 505.3542, 605.381
10	34.047	317.5072	M-H	-8.21	Phytosphingosine	C18H39NO3	316.4966, 352.9604
11	34.395	480.6341	M+Cl	0	Crustecdysone	C27H44O7	516.0871, 75.0456, 99.0812, 115.0761, 141.0934, 145.0868, 291.1613, 301.1825, 445.2586
12	36.535	903.0583	M+Cl	0.53	Trigofoenoside A	C45H74O18	938.5118, 547.3687, 593.3664, 709.4143
13	40.653	196.2429	M-H	6.15	Loliolide	C11H16O3	195.2362, 179.0700, 195.1020
14	41.316	278.3435	M+Cl	-6.05	Dibutyl Phthalate	C16H22O4	313.7946, 55.0548, 147.0078, 167.034, 183.1388
15	42.043	869.0436	M-H	-3.34	Dioscin	C45H72O16	868.0327, 101.0238, 557.3476, 575.3569
16	45.297	274.3978	M-H	5.49	Nandrolone	C18H26O2	273.3914, 147.0822, 173.0955, 239.1447

重楼皂苷VII通过SOHLH1诱导骨肉瘤铁死亡抑制肿瘤进展

Polyphyllin VII inhibits osteosarcoma xenograft growth in mice by inducing ferroptosis via upregulating SOHLH1

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