Clinical significance of miR-34a expression in thyroid diseases – an ¹⁸F-FDG PET-CT study

Figures & data

Table 1 Overview of miR-34a roles in cancers

Cancer type	miR-34a significance	Authors	Reference
Breast carcinoma	Met oncogene is unaffected by miR-34a in nonmetastatic and metastatic breast carcinomas, while in bone metastasis miR-34a correlates with Met, a key player of the bone-metastatic process, expression	Maroni et al	^Citation9
Head and neck squamous cell carcinoma	Forced miR-34a expression induced cell cycle arrest and senescence; genes, including FUT1, AXL and MAP2K1 were identified as the novel targets of miR-34a by qPCR and luciferase assay	Wang et al	^Citation10
Ovarian	Levels of miR-34a are downregulated in ovarian cancer patients. Introduction of miR-34a in ovarian cancer 94 cells resulted in reduced proliferation, motility and invasion
Gastric cancer	Silencing miR-34a activates IGF2BP3, which ranked number 1 of the upregulated genes and associated with poor disease-specific survival	Zhou et al	^Citation11
Osteosarcoma	miR-34a targets DLL1, leading to the ATF2/ATF3/ATF4 signaling pathway, significantly altering and negatively regulating the multi-chemoresistance of osteosarcoma	Pu et al	^Citation12
	miR-34a inhibits growth and metastasis of osteosarcoma cells both in vitro and in vivo through targeting c-Met	Yan et al	^Citation13
Myeloma	Therapeutic potential of synthetic miR-34a against human multiple myeloma was shown partially by repressing BCL-2 and CDK6	Di Martino et al	^Citation14
Hepatocellular carcinoma	miR-34a, by mimicking ST3GAL5, significantly changes HCC malignant behaviors and oncogenicity in in vitro and in vivo assays, acting as tumor suppressor	Cai et al	^Citation15
	miR-34a may inhibit the proliferation of HCC cells and induce their apoptosis via regulating HDAC1 expression	Sun et al	^Citation16
Pancreatic cancer	miR-34a is downregulated in the majority of pancreatic cancers and inhibited the growth of MiaPaCa-2 subcutaneous xenografts	Pramanik et al	^Citation17
Melanoma	Overexpression of miR-34a decreased the invasiveness of melanoma cells, partially involved in P53	Yamazaki et al	^Citation18
Colorectal cancer	miR-34a mediates OXA resistance of CRC by inhibiting macroautophagy via TGF-β/Smad4 pathway	Sun et al	^Citation19
Lung cancer	Hypermethylation of miR-34a promoter was detected in 68.7% lung adenocarcinoma	Daugaard et al	^Citation20
Leukemia	miR-34a promoted cell apoptosis and inhibited autophagy by targeting HMGB1 in acute myeloid leukemia	Liu et al	^Citation21
Renal cell cancer	miR-34a was upregulated in 198 RCC patients and correlated positively with HNF4A	Fritz et al	^Citation22
Cervical cancer	Forced expression of miR-34a inhibited proliferation, migration and invasion by reducing HMGB1 mRNA and protein levels	Chandrasekaran et al	^Citation23
Prostate cancer	Involved in chemoresistance and therapeutic efficacy of PTX in PTC-resistant prostate cancer	Wen et al	^Citation24

Abbreviations: HCC, hepatocellular carcinoma; OXA, oxaliplatin; CRC, colorectal cancer; RCC, renal cell carcinoma; PTX, paclitaxel; PTC, papillary thyroid carcinoma; qPCR, quantitative polymerase chain reaction.

Figure 1 Study flowchart showing the inclusion and exclusion criteria and the entire study design.

Abbreviations: FDG, fluorodeoxyglucose; GLUT1, glucose transporter 1; PET/CT, positron emission tomography/computed tomography; qRT-PCR, quantitative reverse transcription-polymerase chain reaction; SUVmax, maximum standardized uptake value.

Table 2 miR-34a expression in thyroid lesions and clinical characteristics

Clinical characteristics	Patients (n)	miR34a expression (mean ± SD)	P
Age (years)
≤45	41	0.3495 ± 0.02	0.2729
>45	34	0.3759 ± 0.01
Gender
Male	43	0.3551± 0.02	0.5398
Female	32	0.3700 ± 0.02
Tumor stage
I/II	47	0.4111 ± 0.01	<0.0001
III/IV	18	0.2811 ± 0.02
Histopathology
PTC	41	0.4251 ± 0.08	<0.0001
FTC	12	0.3217 ± 0.04
MTC	8	0.3050 ± 0.05
ATC	4	0.2650 ± 0.05
Goiter	10	0.1980 ± 0.03
Lesion numbers
Single	51	0.3629 ± 0.02	0.4865
Multiple	24	0.3442 ± 0.02
Location
Left	38	0.3597 ± 0.11	0.8570
Right	26	0.3600 ± 0.11
Both	11	0.3400 ± 0.12
SUVmax			<0.0001
Low (≤3.63)	27	0.4326 ± 0.02
High (>3.63)	48	0.3098 ± 0.01

Abbreviations: SD, standard deviation; PTC; papillary thyroid carcinoma; FTC, follicular thyroid carcinoma; MTC, medullary thyroid carcinoma; ATC, anaplastic thyroid carcinoma; SUVmax, maximum standardized uptake value.

Figure 2 qRT-PCR results of miR-34a relative expression and the relationship between miR-34a expression and clinical characteristics in enrolled patients. Analysis of miR-34a relative expression by qRT-PCR. The results are shown as 2^−ΔΔCt and normalized to U6. There is no difference in the relative expression of miR-34a between patients of different ages and genders, with different lesion numbers and locations of lesions (data not shown). (A) Patients with higher tumor stage harbored lower miR-34a expression (P < 0.001), while (B) patients with malignant thyroid lesions expressed higher miR-34a than those with goiter (P < 0.0001). We also analyzed the miR-34a expression among different malignant pathology types and found that there is no significant difference among FTC, MTC and ATC. (C) However, tendency shows that ATC harbored lower miR-34a compared to other groups. (D) miR-34a expression was significantly higher in patients with low SUVmax (SUVmax ≤ 3.63) than in those with higher SUVmax (SUVmax >3.63, P < 0.0001). ***P < 0.001.

Abbreviations: FDG, fluorodeoxyglucose; FTC, follicular thyroid carcinoma; MTC, medullary thyroid carcinoma; ATC, anaplastic thyroid carcinoma; SUVmax, maximum standardized uptake value; PTC, papillary thyroid carcinoma.

Figure 3 Graphs show the relationship between miR-34a expression and SUVmax in patients with thyroid diseases. (A) There was an inverse relationship between miR-34a expression and SUVmax in patients with thyroid diseases (Spearman correlation coefficient = −0.553, P < 0.0001). (B) ROC curve analysis of SUVmax to predict miR-34a expression. The miR-34a expression was divided into 2 different groups, named as miR-34a-L (whose miR-34a expression is lower than or equal to 0.36) and miR-34a-H (miR-34a expression is higher than 0.36). With an SUVmax of 4.3 as the threshold, sensitivity and specificity of the prediction of miR-34a expression (low or high) were 70% and 94.3%, respectively. The area under the ROC curve was 0.843 (95% confidence interval: 0.749, 0.936; P = 0.001).

Abbreviations: SUVmax, maximum standardized uptake value; ROC, receiver operating characteristic.

Figure 4 Inhibiting miR-34a expression increased GLUT1 expression. Graphs show the influence of miR-34a inhibition on glucose transporters in cultured TPC-1 cells. miR-34a may decrease ¹⁸F-FDG uptake via targeting GLUT1 expression. Data are mean ± standard deviation. (A) miR-34a inhibitor successfully downregulated the miR-34a expression (P < 0.001), while there is no significant statistical difference between TPC-1 and NC group (P > 0.05). (B and C) Influence of miR-34a inhibition on expression of GLUT1 mRNA and protein in TPC-1 cells. miR-34a inhibition significantly increased expression of GLUT1 mRNA and protein (both P < 0.001). ***P < 0.001.

Abbreviations: GLUT1, glucose transporter 1; FDG, fluorodeoxyglucose; NC, negative control.

Figure 5 Models show the possible metabolic status in thyroid cancer with different miR-34a expression. miR-34a expression in ATC was lower than that in differentiated thyroid carcinoma, which cannot inhibit the GLUT1 expression, leading to increased FDG avidity. The glycolysis pathway, including ¹⁸F-FDG uptake, was upregulated in patients with ATC. Thus, SUVmax was significantly higher in patients with ATC.

Abbreviations: ATC, anaplastic thyroid carcinoma; GLUT1, glucose transporter 1; FDG, fluorodeoxyglucose; SUVmax, maximum standardized uptake value; miRNA, microRNA.

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