Research on the effect of tomato derived nanovesicles on oral squamous cell carcinoma

doi:10.11958/20252768

Abstract

Abstract:

Objective To explore the effect of tomato derived nanovesicles (TDNVs) on the malignant biological behavior of oral squamous cell carcinoma (OSCC) and its potential molecular mechanism. Methods TDNVs in tomato fruits were extracted and purified by optimizing ultracentrifugation and sucrose density gradient centrifugation. The morphology and particle size were identified by transmission electron microscopy (TEM) and nanoparticle tracking analysis (NTA) experiments. Coomassie Brilliant Blue staining was employed to detect the TDNVs protein content. The effects of TDNVs on the proliferation, cell cycle and apoptosis abilities of WSU-HN6 and SCC-15 cells were detected by cell counting kit 8 (CCK-8) and flow cytometry, respectively. The expression of genes related to cell cycle and apoptosis was detected by Real-time PCR and Western blot experiment. Results TDNVs were successfully isolated and identified, and the average particle size of TDNVs was 167.8 nm. The particle concentration is approximately 1.3×10¹¹ particles/mL. In vitro experiments demonstrated that TDNVs could significantly inhibit the proliferation ability of OSCC cells (P<0.05), induced cell cycle arrest at the G₂/M phase and promoted cell apoptosis (P<0.05). Meanwhile, TDNVs treatment led to a decrease in the mRNA expression levels of cyclin-dependent kinase 1 (CDK1), Cyclin B1 and B-lymphoblastoma-2 gene (Bcl-2) in OSCC cells (P<0.05). The mRNA expression level of Bcl-2 associated X protein (Bax) increased (P<0.05). Conclusion TDNVs can inhibit the malignant progression of OSCC cells by inducing cell cycle arrest and promoting apoptosis.

Key words: mouth neoplasms, carcinoma, squamous cell, lycopersicon esculentum, extracellular vesicles, cell proliferation, apoptosis, cell cycle

CLC Number:

R739.81

Figures/Tables 13

References 20

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基因	引物序列（5′→3′）	产物大小/bp
GAPDH	上游：TCAGCAATGCCTCCTGCAC	117
	下游：TCTGGGTGGCAGTGATGGC	117
CDK1	上游：AGTCTACGGGCTACCCGATT	706
	下游：CCACTCTGCCCTAGGCTTTC	706
Cyclin B1	上游：TGGTGAATGGACTGTCAAGAACA	117
	下游：TACACCTTTGCCACAGCCTT	117
Bcl-2	上游：AAAAATACAACATCACAGAGGAAGT	334
	下游：TCCCGGTTATCGTACCCTGT	334
Bax	上游：TGATGGACGGGTCCGGG	422
	下游：TGTCCAGCCCATGATGGTTC	422

基因	引物序列（5′→3′）	产物大小/bp
GAPDH	上游：TCAGCAATGCCTCCTGCAC	117
	下游：TCTGGGTGGCAGTGATGGC	117
CDK1	上游：AGTCTACGGGCTACCCGATT	706
	下游：CCACTCTGCCCTAGGCTTTC	706
Cyclin B1	上游：TGGTGAATGGACTGTCAAGAACA	117
	下游：TACACCTTTGCCACAGCCTT	117
Bcl-2	上游：AAAAATACAACATCACAGAGGAAGT	334
	下游：TCCCGGTTATCGTACCCTGT	334
Bax	上游：TGATGGACGGGTCCGGG	422
	下游：TGTCCAGCCCATGATGGTTC	422

组别	WSU-HN6			SCC-15
组别	G₁	S	G₂/M	G₁	S	G₂/M
Blank组	65.38±2.98	16.45±2.15	18.16±1.65	56.30±3.49	28.10±1.81	15.60±2.01
PBS组	63.09±3.97	16.09±3.54	20.82±1.42	55.52±2.99	26.80±3.07	17.69±2.32
5 mg/L组	34.64±2.22^ab	9.70±0.66^ab	55.66±2.06^ab	40.17±1.69^ab	25.57±2.20	34.26±0.56^ab
F	89.061^＊＊	7.376^＊	439.508^＊＊	31.068^＊＊	0.817	96.625^＊＊

组别	WSU-HN6			SCC-15
组别	G₁	S	G₂/M	G₁	S	G₂/M
Blank组	65.38±2.98	16.45±2.15	18.16±1.65	56.30±3.49	28.10±1.81	15.60±2.01
PBS组	63.09±3.97	16.09±3.54	20.82±1.42	55.52±2.99	26.80±3.07	17.69±2.32
5 mg/L组	34.64±2.22^ab	9.70±0.66^ab	55.66±2.06^ab	40.17±1.69^ab	25.57±2.20	34.26±0.56^ab
F	89.061^＊＊	7.376^＊	439.508^＊＊	31.068^＊＊	0.817	96.625^＊＊

组别	WSU-HN6		SCC-15
组别	CDK1	Cyclin B1	CDK1	Cyclin B1
Blank组	1.00±0.06	1.00±0.02	1.00±0.04	1.00±0.01
PBS组	0.97±0.08	1.02±0.16	1.06±0.07	1.04±0.23
5 mg/L组	0.61±0.02^ab	0.45±0.04^ab	0.73±0.03^ab	0.64±0.04^ab
F	82.974^＊＊	40.112^＊＊	36.553^＊＊	8.088^＊