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中华细胞与干细胞杂志(电子版) ›› 2022, Vol. 12 ›› Issue (04) : 200 -205. doi: 10.3877/cma.j.issn.2095-1221.2022.04.002

论著

柴胡皂苷D对甲状腺乳头状癌细胞TPC-1生物学行为的影响
周辰杰1,(), 莫雪林1, 韩炜蔚1   
  1. 1. 215000 苏州,江苏省苏州市南京医科大学附属苏州科技城医院药学部
  • 收稿日期:2022-03-30 出版日期:2022-08-01
  • 通信作者: 周辰杰
  • 基金资助:
    南京医科大学科技发展基金(NMUB2020249)

Effects of saikosaponin D on the biological behavior of papillary thyroid carcinoma TPC-1

Chenjie Zhou1,(), Xuelin Mo1, Weiwei Han1   

  1. 1. Department of Pharmacy, Suzhou Science and Technology City Hospital Affiliated to Nanjing Medical University, Suzhou 215000, China
  • Received:2022-03-30 Published:2022-08-01
  • Corresponding author: Chenjie Zhou
引用本文:

周辰杰, 莫雪林, 韩炜蔚. 柴胡皂苷D对甲状腺乳头状癌细胞TPC-1生物学行为的影响[J/OL]. 中华细胞与干细胞杂志(电子版), 2022, 12(04): 200-205.

Chenjie Zhou, Xuelin Mo, Weiwei Han. Effects of saikosaponin D on the biological behavior of papillary thyroid carcinoma TPC-1[J/OL]. Chinese Journal of Cell and Stem Cell(Electronic Edition), 2022, 12(04): 200-205.

目的

探讨柴胡皂苷D对体外甲状腺乳头状癌(PTC)细胞TPC-1增殖、侵袭、迁移、活性氧和凋亡的影响。

方法

不同浓度(0、5、10、20 μmol/L)柴胡皂苷D干预TPC-1细胞,CCK-8法检测细胞增殖率;Annexin-V-FITC/PI双染试剂盒检测细胞凋亡率;活性氧试剂盒检测细胞活性氧;Transwell小室法检测细胞侵袭、迁移能力;Western blot法检测凋亡蛋白剪切的半胱天冬酶3 (cleaved caspase 3)、B淋巴细胞瘤-2基因(Bcl-2)、Bcl2相关X蛋白(Bax)和侵袭迁移相关蛋白E钙黏蛋白(E-cadherin)和N钙黏蛋白(N-cadherin)及Wnt/β连环蛋白(β-catenin)信号通路相关蛋白Wnt3a和β-catenin的表达量。多组间比较采用单因素方差分析,组间两两比较采用LSD-t检验。

结果

与0 μmol/L柴胡皂苷D干预相比,(5、10、20 μmol/L)柴胡皂苷D干预TPC-1细胞存活率[24 h:(88.07±0.75)%、(76.06±0.56)%、(65.80± 1.66)%比(100.00±1.00)%;48 h:(72.40±0.35)%、(52.26± 0.22)%、(41.30±0.17)%比(100.00± 1.53)%;72 h:(56.50±0.22)%、(40.26±0.21)%、(31.10± 0.14)%比(100.00±0.14)%]、细胞侵袭率[(65.43±1.32)%、(49.46±0.31)%、(22.48±0.85)%比(100.00±1.25)%]、迁移率[(67.45±0.96)%、(52.42±0.39)%、(42.12±1.10)%比(100.00±2.00)%]、Bcl-2蛋白表达量(0.51±0.02、0.31±0.02、0.22±0.02比0.62±0.01)、Wnt3a表达量(0.56±0.06、0.37±0.01、0.26±0.02比0.72±0.03)、β-catenin表达量(0.59±0.02、0.40±0.03、0.14±0.03比0.82±0.03)和N-cadherin表达量(0.49±0.05、0.34 ±0.03、0.14± 0.03比0.62±0.05)均降低,细胞活性氧含量增加,凋亡率[(12.36±1.32)%、(23.15±2.27)%、(36.65±3.22)%比(7.32±1.25)%]、Bax蛋白表达量(0.45±0.02、0.51±0.01、0.79±0.02比0.32±0.01)、cleaved caspase-3蛋白表达量(0.26±0.02、0.37±0.02、0.40±0.02比0.13±0.01)和E-cadherin表达量(0.20±0.02、0.37±0.02、0.46±0.05比0.12±0.01)均增加(P均< 0.05)。

结论

柴胡皂苷D能够抑制PTC细胞的生物学行为。

Objective

To explore the effect of saikosaponin D on the proliferation, invasion, migration, reactive oxygen species and apoptosis of TPC-1 papillary thyroid carcinoma (PTC) cells in vitro.

Methods

The cells were treated with 0、5、10、20 μmol/L saikosaponin D. The cell proliferation rate was detected by CCK-8 assay. Annexin V conjugate (Annexin V-FITC) and propidium iodide (PI) double staining kit was used to detect cell apoptosis rate. Reactive oxygen detection kit was used to detect cell reactive oxygen species. Transwell chamber method was used to detect cell invasion and migration ability. Western blot was used to detect the expression of apoptosis proteins including cleaved caspase-3, B-cell lymphoma-2 (Bcl-2) , Bcl2 associated X protein (Bax) , invasion and migration related proteins E-cadherin, N-cadherin and Wnt/β-catenin signaling pathway related proteins Wnt3a and β-catenin. At the same time, one-way ANOVA was used to compare multiple groups and the LSD-t test was used for pairwise comparison between groups.

Results

Compared with 0 μmol/L saikosaponin D, 5、10、20 μmol/L saikosaponin D significantly decreased survival rate (24 h: 88.07±0.75, 76.06±0.56, 65.80±1.66 vs 100.00±1.00. 48 h: 72.40±0.35, 52.26±0.22, 41.30±0.17 vs 100.00±1.53. 72 h: 56.50±0.22, 40.26±0.21, 31.10±0.14 vs 100.00±0.14) , cell invasion rate [ (65.43±1.32) %, (49.46±0.31) %, (22.48±0.85) % vs (100.00±1.25) %], mobility[ (67.45±0.96) %, (52.42±0.39) %, (42.12± 1.10) % vs (100.00±2.00) %], Bcl-2 protein expression (0.51±0.02, 0.31±0.02, 0.22±0.02 vs 0.62±0.01) , Wnt3a expression (0.56±0.06, 0.37±0.01, 0.26±0.02 vs 0.72±0.03) , β-catenin expression (0.59±0.02, 0.40±0.03, 0.14±0.03 vs 0.82±0.03) , N-cadherin expression (0.49±0.05, 0.34±0.03, 0.14±0.03 vs 0.62±0.05) in TPC-1 cells. Compared with 0 μmol/L saikosaponin D, 5、10、20 μmol/L saikosaponin D significantly increased the content of reactive oxygen species in cells and the apoptosis rate[ (12.36±1.32) %, (23.15±2.27) %, (36.65±1.32) %vs (7.32±1.25) %], Bax protein expression (0.45±0.02, 0.51±0.01, 0.79±0.02 vs 0.32±0.01) , cleaved caspase-3 protein expression (0.26±0.02, 0.37±0.02, 0.40±0.02 vs 0.13±0.01) and E-cadherin expression (0.20±0.02, 0.37±0.02, 0.46±0.05 vs 0.12±0.01) (all P < 0.05) .

Conclusion

Saikosaponin D can inhibit the biological behavior of PTC cells.

表1 柴胡皂苷D作用于TPC-1细胞后细胞凋亡率及cleaved caspase-3、Bax、Bcl-2蛋白相对表达量(±s
图1 柴胡皂苷D对TPC-1细胞活性氧与凋亡的影响注:a ~ d图为荧光显微镜下观察0、5、10、20 μmol/L柴胡皂苷D干预TPC-1细胞的活性氧,绿色荧光越强表明活性氧含量越高(×100);e ~ h图分别为0、5、10、20 μmol/L柴胡皂苷D干预TPC-1细胞的凋亡率
图2 Western blot检测cleaved caspase-3、Bax和Bcl-2蛋白相对表达量
表2 柴胡皂苷D作用于TPC-1细胞后侵袭、迁移能力与Wnt3a、β-catenin、E-cadherin、N-cadherin相对表达量(±s
图3 Transwell小室法检测柴胡皂苷D对TPC-1细胞侵袭和迁移的影响(结晶紫染色,×100)注:a ~ d图为倒置荧光显微镜下观察0、5、10、20 μmol/L柴胡皂苷D干预TPC-1细胞的侵袭结果;e ~ h图分别为倒置荧光显微镜下观察0、5、10、20 μmol/L柴胡皂苷D干预TPC-1细胞的迁移结果
图4 Western blot检测Wnt3a、β-catenin、E-cadherin和N-cadherin的相对表达量
1
Liu Y, Zhang H, Wang H, et al. Long non-coding RNA DUXAP8 promotes the cell proliferation, migration, and invasion of papillary thyroid carcinoma via miR-223-3p mediated regulation of CXCR4[J]. Bioengineered, 2021, 12(1):496-506.
2
Du YL, Liang Y, Cao Y, et al. LncRNA XIST promotes migration and invasion of papillary thyroid cancer cell by modulating MiR-101-3p/CLDN1 Axis[J]. Biochem Genet, 2021, 59(2):437-452.
3
Li Y, Zeng QG, Qiu JL, et al. Propofol suppresses migration, invasion, and epithelial-mesenchymal transition in papillary thyroid carcinoma cells by regulating miR-122 expression[J]. Eur Rev Med Pharmacol Sci, 2020, 24(9):5101-5110.
4
Li C, Xue HG, Feng LJ, et al. The effect of saikosaponin D on doxorubicin pharmacokinetics and its MDR reversal in MCF-7/adr cell xenografts[J]. Eur Rev Med Pharmacol Sci, 2017, 21(19):4437-4445.
5
Li Y, Cai T, Zhang W, et al. Effects of Saikosaponin D on apoptosis in human U87 glioblastoma cells[J]. Mol Med Rep, 2017, 16(2):1459-1464.
6
Hsu YL, Kuo PL, Lin CC. The proliferative inhibition and apoptotic mechanism of saikosaponin D in human non-small cell lung cancer A549 cells[J]. Life Sci, 2004, 75(10):1231-1242.
7
Ma Q, Gao FF, He X, et al. Antitumor effects of saikosaponin b2 on breast cancer cell proliferation and migration[J]. Mol Med Rep, 2019, 20(2):1943-1951.
8
Yao M, Yang J, Cao L, et al. Saikosaponind inhibits proliferation of DU145 human prostate cancer cells by inducing apoptosis and arresting the cell cycle at G0/G1 phase[J]. Mol Med Rep, 2014, 10(1):365-372.
9
Ma X, Dang C, Kang H, et al. Saikosaponin-D reduces cisplatin-induced nephrotoxicity by repressing ROS-mediated activation of MAPK and NF-κB signalling pathways[J]. Int Immunopharmacol, 2015, 28(1):399-408.
10
Gao T, Zhao P, Yu X, et al. Use of saikosaponin D and JNK inhibitor SP600125, alone or in combination, inhibits malignant properties of human osteosarcoma U2 cells[J]. Am J Transl Res, 2019, 11(4):2070-2080.
11
Tang JC, Long F, Zhao J, et al. The effects and mechanisms by which saikosaponin-d enhances the sensitivity of human non-small cell lung cancer cells to gefitinib[J]. J Cancer, 2019, 10(26):6666-6672.
12
Zhong D, Zhang HJ, Jiang YD, et al. Saikosaponin-d: A potential chemotherapeutics in castration resistant prostate cancer by suppressing cancer metastases and cancer stem cell phenotypes[J]. Biochem Biophys Res Commun, 2016, 474(4):722-729.
13
Tian YD, Lin S, Yang PT, et al. Saikosaponin-d increases the radiosensitivity of hepatoma cells by adjusting cell autophagy[J]. J Cancer, 2019, 10(20):4947-4953.
14
Cai C, Zhang H, Ou Y, et al. Saikosaponin-d suppresses cell growth in renal cell carcinoma through EGFR/p38 signaling pathway[J]. Neoplasma, 2017, 64(4):518-525.
15
He S, Lu G, Hou H, et al. Saikosaponind suppresses the expression of cyclooxygenase2 through the phosphosignal transducer and activator of transcription 3/hypoxiainducible factor1α pathway in hepatocellular carcinoma cells[J]. Mol Med Rep, 2014, 10(5):2556-2562.
16
Zhao L, Li J, Sun Z, et al. Saikosaponin D inhibits proliferation of human osteosarcoma cells via the p53 signaling pathway[J]. Exp Ther Med, 2019, 17(1):488-494.
17
Sun K, Du Y, Hou Y, et al. Saikosaponin D exhibits anti-leukemic activity by targeting FTO/mA signaling[J]. Theranostics, 2021, 11(12):5831-5846.
18
Tsuyoshi H, Wong VKW, Han Y, et al. Saikosaponin-d, a calcium mobilizing agent, sensitizes chemoresistant ovarian cancer cells to cisplatin-induced apoptosis by facilitating mitochondrial fission and G2/M arrest[J]. Oncotarget, 2017, 8(59):99825-99840.
19
Zhou P, Shi W, He X, et al. Saikosaponin D: review on the antitumour effects, toxicity and pharmacokinetics[J]. Pharm Biol, 2021, 59(1):1480-1489.
20
Chen X, Liu C, Zhao R, et al. Synergetic and antagonistic molecular effects mediated by the feedback loop of p53 and JNK between saikosaponin D and SP600125 on lung cancer A549 cells[J]. Mol Pharm, 2018, 15(11):4974-4984.
21
Liu RY, Li JP. Saikosaponin-d inhibits proliferation of human undifferentiated thyroid carcinoma cells through induction of apoptosis and cell cycle arrest[J] .Eur Rev Med Pharmacol Sci, 2014, 18(17):2435-2443.
22
Lv Y, Hou X, Zhang Q, et al. Untargeted metabolomics study of the in vitro anti-hepatoma effect of saikosaponin D in combination with NRP-1 knockdown[J]. Molecules, 2019, 24(7):1423. doi:10.3390/molecules24071423.
23
Wang B, Min W, Lin S, et al. Saikosaponin-d increases radiation-induced apoptosis of hepatoma cells by promoting autophagy via inhibiting mTOR phosphorylation[J]. Int J Med Sci, 2021, 18(6):1465-1473.
24
Wu S, Chen W, Liu K, et al. Saikosaponin D inhibits proliferation and induces apoptosis of non-small cell lung cancer cells by inhibiting the STAT3 pathway[J] J Int Med Res, 2020, 48(9):300060520937163. doi: 10.1177/0300060520937163.
25
Fu R, Zhang L, Li Y, et al. Saikosaponin D inhibits autophagosome lysosome fusion and induces autophagy independent apoptosis in MDA MB 231 breast cancer cells[J] Mol Med Rep, 2020, 22(2):1026-1034.
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