Influence of MMP12 Single Nucleotide Polymorphism rs586701 on the Prognosis of Primary Lung Cancer in Chinese Han Chinese

doi:10.21203/rs.3.rs-4711599/v1

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Research Article

Influence of MMP12 Single Nucleotide Polymorphism rs586701 on the Prognosis of Primary Lung Cancer in Chinese Han Chinese

https://doi.org/10.21203/rs.3.rs-4711599/v1

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Purpose

Lung cancer is now one of the most common cancers in the world, with a high mortality rate and poor prognosis. Predicting the prognosis of lung cancer patients and using this information to develop treatment strategies and interventions is important for prolonging patient survival. A stratified analysis was conducted on Han Chinese primary lung cancer patients in the Chinese population, with the objective of investigating the relationship between matrix metalloproteinase 12(MMP12) gene polymorphism rs586701 and the prognosis of lung cancer patients.

Methods

A total of 888 Han Chinese primary lung cancer patients (exclusive of minors) were recruited between January and November 2009 (10 months) and provided written informed consent. The study included subjects from Changhai Hospital of the Naval Medical University (Second Military Medical University) and Taizhou Institute of Health Sciences of Fudan University. A total of 49 subjects were excluded due to incomplete data collection for various reasons. Blood samples were collected from 839 patients diagnosed with lung cancer, and genomic DNA was extracted for genotyping using SNPscan technology. In order to adjust for multiple factors, the data was stratified by age, sex, smoking status, family history of malignancy, TNM stage, and lung cancer tissue type. The association between lung cancer prognosis and genotype was then analyzed using a multivariate Cox proportional risk model.

Results

A polymorphism in the MMP12 gene, specifically the T > G variant at position 586701, has been associated with a reduction in survival time in lung cancer patients. In the overall sample, patients with the TG genotype (TG vs TT, adjusted HR = 1.21, 95% CI: 1.01–1.44, P = 0.035) exhibited a shorter survival time and a worse prognosis. Stratified analyses showed that among male lung cancer patients, lung cancer patients younger than 60 years old, and smoking lung cancer patients, patients with the TG genotype had a lower survival time than patients with the TT genotype (adjusted risk ratio HR = 1.24, 95% CI: 1.01–1.52, P = 0.04; HR = 1.58, 95% CI: 1.18–2.12, P = 0.002; HR= (1.30, 95% CI: 1.06–1.61, P = 0.013). In patients with SCC and NSCLC, survival time was shorter in patients with the TG genotype (TG vs TT, adjusted risk ratio HR = 1.48, 95% CI: 1.10–2.01, P = 0.010; HR = 1.21, 95% CI: 1.01–1.46, P = 0.038). In the designed genetic model, the dominant genotype TG + GG was associated with a shorter survival time and worse prognosis among lung cancer patients aged less than 60 years (TG + GG vs TT, adjusted risk ratio HR = 1.43, 95% CI: 1.07–1.90, P = 0.014).

Conclusion

The MMP12 polymorphism rs586701 may be associated with the prognosis of lung cancer. Specifically, the MMP12 polymorphism rs586701 T > G has been found to lead to a worse prognosis.

MMP12

single nucleotide polymorphism

rs586701

lung cancer

prognosis

Lung cancer represents the leading cause of cancer-related mortality on a global scale [1]. Lung cancer has the highest incidence and mortality rate in China[2]. Although early diagnosis and treatment of lung cancer are improving, the clinical prognosis remains poor, with a five-year survival rate of less than 20%[3]. The establishment of accurate prognostic indicators has been demonstrated to improve treatment outcomes and prolong patient survival in lung cancer[4]. Epidemiological studies have demonstrated that age, male sex, smoking history, small cell lung cancer, and advanced lung cancer (stage III or IV) are associated with a poor prognosis for lung cancer[5]. Furthermore, an increasing number of studies have indicated that individual genetic factors may be of significant importance in the prognosis of lung cancer[6], Single nucleotide polymorphisms (SNPs), which are common genetic variants that can affect gene expression and the prognosis of lung cancer patients, are a notable example of this [7–10].

The family of matrix metalloproteinases (MMPs) belongs to the family of zinc-dependent endopeptidases, a class of proteases that degrade the extracellular matrix and basement membrane barrier[11, 12]. In addition to their involvement in multiple stages of cancer development, they also play a significant role in tumor invasion and metastasis[13]. A total of 20 or more distinct types of matrix metalloproteinases (MMPs) have been identified. MMPs can be classified according to their structural features and sensitivity to substrates. The collagenases (MMP-1, MMP-8, MMP-13, and MMP-18), gelatinases (MMP-2 and MMP-9), and matrix proteins (MMP-3, MMP-10, and so on) are examples of this classification. MMP-11, matrices (MMP-7 and MMP-26), GPI anchors (MMP-17 and MMP-25), membrane-type MMPs (MMP-14, MMP-15, MMP-16, and MMP-24), and so on. Membrane-type MMPs (MMP-14, MMP-15, MMP-16, and MMP-24) and other[14]. MMP12 is located on chromosome 11q22.3 in the cluster of genes encoding matrix metalloproteinases and encodes a member of the M10 family of matrix metalloproteinase (MMP) peptidases. Proteins in this family are involved in the breakdown of extracellular matrices during normal physiological processes, including embryonic development, reproduction, and tissue remodeling. They are also involved in disease processes, such as arthritis and metastasis. The encoded procollagen undergoes proteolytic processing to produce mature proteases that degrade soluble and insoluble elastin.。

Molecular epidemiological studies have demonstrated that genetic polymorphisms in matrix metalloproteinase (MMP) are associated with susceptibility and prognosis in lung cancer[15–17], adenocarcinoma[18, 19], colorectal cancer[20, 21], cervical cancer[22, 23] and other cancers[24, 25]. The MMP9-1562C/T polymorphism and the MMP13-77G/A polymorphism have been linked to an increased risk of developing NSCLC [26]. Certain MMP3 promoter polymorphisms have been associated with an elevated susceptibility to NSCLC and an increased risk of lymph node metastasis[27]. It has been demonstrated that polymorphisms in the MMP2, MMP9, and MMP12 genes, specifically rs243864, rs3918242, and rs652438, respectively, influence the transcriptional activity and expression level of the encoded protein[28–30], which in turn affects prognosis. rs586701 is situated within the gene region between MMP3 and MMP12. However, the relationship between the MMP12 polymorphism rs586701 and lung cancer prognosis has not yet been elucidated. Therefore, we conducted a study to investigate the association between the MMP12 polymorphism rs586701 and the prognosis of lung cancer. To do this, we stratified the lung cancer population by age, sex, smoking status, family history, lung cancer stage, and tissue type. Additionally, we collected pre-treatment peripheral blood samples from lung cancer patients and performed genotyping and follow-up.

2.1 Ethics Statement

The entire study design protocol was approved by the Ethics Committee of the School of Life Sciences, Fudan University. Prior to the collection of blood samples, each subject was provided with a standard informed consent form, which clearly explained the purpose and procedures of the study. All data were recorded anonymously.

2.2 Subjects of study and data collection

A total of 888 patients with primary lung cancer were admitted to the Naval Military Medical University (NMMU) and the Institute of Health Sciences, Fudan University Taizhou (IHSFT) between January and November 2009. Of these, 536 cases were from Changhai Hospital affiliated to the NMMU, and 352 cases were from the Institute of Health Sciences, Fudan University Taizhou. The follow-up period commenced at the time of inclusion in the study and concluded at the end of 2019. During this period, 49 patients were excluded due to incomplete clinical data. The remaining 839 patients' data were collected and subsequently analyzed. The study population consisted of Han Chinese individuals. Inclusion criteria included histological diagnosis of primary lung cancer and no history of malignant tumors in other organs. There were no restrictions on age or gender, and cancer stage was not a limiting factor. Histological diagnosis of lung cancer was based on World Health Organization criteria, and lung cancer stage was determined according to the 8th edition of TNM staging of lung cancer, with confirmation by two independent pathologists. The initial stages (I and II) were considered to be of an early nature, while the subsequent stages (III and IV) were regarded as being of a later chronology. The clinical data pertaining to the patients was obtained from their medical records, while the follow-up information was derived from telephone interviews.

2.3 SNP genotyping

Venous blood samples were collected from all enrolled patients prior to the commencement of treatment. Genomic DNA was extracted using the Qiagen Blood DNA Extraction Kit (Qiagen, Hilden, Germany), and the resulting DNA samples were amplified by polymerase chain reaction (PCR),and genotyping was conducted using the 2×48-plex SNPscan TM kit (order no. G0104; Genentech, Shanghai, China) that based on double ligation and multiplex fluorescence PCR. SNPscan is a proprietary multiple SNP genotyping system that permits the simultaneous genotyping of 48, 96, 144, or 192 SNPs per sample in a single tube or sample. SNPscan employs a highly specific linkage reaction to distinguish between alleles. The genotyping quality was determined using a detailed procedure that included a successful detection rate of more than 95%, the identification of duplicate genotypes, the use of internal positive control samples, and Hardy-Weinberg equilibrium (HWE) testing. Furthermore, the laboratory personnel responsible for the genotyping analysis were unaware of the patient's clinical information.

2.4 Statistical analysis

Prior to performing the association analysis, the Hardy-Weinberg equilibrium (HWE) test was conducted on rs586701 within the study population using the Pearson chi-square test. Overall survival (OS) was calculated from the date of sample collection to the date of death from any cause or the date of the last follow-up visit. Median survival time (MST) was estimated by the Kaplan-Meier method, and differences between groups were tested by the log-rank test. Univariate and multivariate Cox regression analyses were employed to estimate hazard ratios (HR) and their 95% confidence intervals (CI), adjusted for age and sex. A stratified analysis was conducted according to age, sex, smoking status, family history of malignancy, TNM stage, and histologic type of lung cancer. The relationships between single nucleotide polymorphisms (SNPs) and clinical outcomes in diverse populations of lung cancer patients were evaluated using four SNP genetic models: allelic, genotypic, dominant, and recessive. These models were stratified according to age, gender, smoking status, family history, lung cancer stage, and tissue type. All tests were two-sided and statistically significant at the 0.05 level. All statistical analyses were performed using R version 3.6.2.

3.1 Demographic and clinical characteristics and prognostic analysis.

Table 1 presents a summary of the demographic characteristics of the 839 lung cancer patients. During the follow-up period, 668 patients (79.6%) died, 103 patients (12.3%) survived for more than five years, and 68 patients (8.1%) were lost to follow-up. The mean survival time (MST) for all patients was 36.73 months. Of all patients, 229 (27.3%) were female and 610 (72.7%) were male. In total, 315 (37.5%) were below 60 years of age and 524 (62.5%) were above 60 years of age. A total of 237 patients (28.2%) had never smoked, while 582 patients (69.4%) had a history of smoking. Of the patients, 537 (64%) had no family history of malignancy, while 302 (36%) had a family history of malignancy. The histological diagnoses were as follows: 367 patients (43.7%) were diagnosed with adenocarcinoma (ADC), 282 patients (33.6%) with squamous cell carcinoma (SCC), 72 patients (8.6%) with small cell lung cancer (SCLC), and 118 patients (14.1%) with other types of cancer. The TNM staging system revealed that 154 cases (18.4%) exhibited stage I and II lung cancer, while 625 cases (74.5%) exhibited stage III and IV lung cancer.

Table 1

Characteristic distribution in Chinese patients with lung cancer and prognosis analysis.
Variables	N(%)	MST^#	P
Total	839	36.73
Sex			0.01
Female	229(27.3%)	40.17
Male	610(72.7%)	34.27
Age			0.003
<60	315(37.5%)	40.87
≥60	524(62.5%)	33.20
Smoking stage
Nonsmoker	237(28.2%)	41.03
Smoker	582(69.4%)	33.90
Unknown	20(2.4%)	67
Family History			0.462
Yes	302(36%)	33.63
No	537(64%)	38.03
Subtype			0.211
ADC	367(43.7%)	38.80
SCC	282(33.6%)	33.63
SCLC	72(8.6%)	33.90
Others	118(14.1%)	36.20
TNM stage			< 0.001
StageⅠ+Ⅱ	154(18.4%)	113.93
StageⅢ+Ⅳ	625(74.5%)	29.4
Unknown	60(7.1%)	66.43

The log-rank test demonstrated statistically significant differences in median survival time (MST) with respect to gender, age, smoking status, and clinical stage (log-rank P < 0.05). The mean survival time (MST) was significantly longer in women, patients aged < 60 years, nonsmokers, and patients with early-stage tumors than in men (MST:40.17 M vs. 34.27 M; P = 0.01), and in patients aged ≥ 60 years (MST:40.87 M vs. 33.2 M). The mean survival time was 33.2 months in smokers (P < 0.001), 41.03 months in patients with advanced tumors (P < 0.001), and 113.93 months in patients who were smokers (P < 0.001).

3.2 Correlation between MMP12 polymorphisms and prognosis of lung cancer.

A total of 817 genotypes were identified, with a detection rate of 97.38%. Of these, 607 were TT genotypes, 208 were TG genotypes, 24 were GG genotypes, and the frequency of the MMP12 rs The 586701 genotypes were consistent with the Hardy-Weinberg equilibrium (P = 0.2051), indicating that the lung cancer patients participating in the study were in a state of genetic equilibrium and that the data obtained from the lung cancer patients studied were credible. The frequencies of rs586701 alleles T and G were 84.73% (1132/1336) and 15.27% (204/1336) in deceased patients and 84.80% (290/342) and 15.20% (52/342) in surviving patients, respectively. Univariate Cox regression analysis revealed that in the overall sample, patients with the TG genotype exhibited a poorer prognosis than patients with the TT genotype (TG vs TT, HR = 1.21, 95% CI:1.01–1.44, P = 0.034; Table 2). Kaplan-Meier survival curves demonstrated that patients with the TG genotype exhibited a shorter median survival time (MST) compared to patients with the TT genotype (MST:38.5 M vs 29.53 M). This difference was statistically significant (Log-rank P = 0.035; Fig. 1).

Table 2

Association between *MMP12* gene polymorphism rs586701 and prognosis of Chinese lung cancer patients
Model	Death/survive	MST	HR(95% Cl)^*	P	HR^a(95%Cl)	P^a
Allele
T(ref)	1132/290	37.13	1		1
G	204/52	32.7	1.03 (0.89–1.20)	0.698	1.03 (0.89–1.20)	0.671
Genotype
T/T(ref)	479/128	38.5	1		1
T/G	174/34	29.53	1.21 (1.01–1.44)	0.034	1.21 (1.01–1.44)	0.035
G/G	15/9	67.33	0.60 (0.35–1.02)	0.059	0.61 (0.36–1.04)	0.067
Dominate
T/T(ref)	479/128	38.5	1		1
T/G + G/G	189/43	31.43	1.12 (0.95–1.33)	0.181	1.12 (0.95–1.33)	0.179
Recessive
T/T + T/G(ref)	653/162	36.03	1		1
G/G	15/9	67.33	0.57 (0.34–0.97)	0.038	0.58 (0.34–0.99)	0.045

3.3 Stratified analysis of the association between MMP12 polymorphisms and lung cancer prognosis.

A stratified analysis was conducted to investigate the association between the MMP12 polymorphism rs586701 and the prognosis of lung cancer patients. Stratified analyses showed (Table 3) that among male lung cancer patients, lung cancer patients younger than 60 years old, and smoking lung cancer patients, patients with the TG genotype had a shorter survival time than patients with the TT genotype (adjusted risk ratios HR = 1.24, 95% CI: 1.01–1.52, P = 0.04; HR = 1.58, 95% CI: 1.18–2.12, P = 0.002 ; HR = 1.30, 95% CI: 1.06–1.61, P = 0.013); in patients with SCC, survival time was shorter in patients with NSCLC, and in patients with the TG genotype (TG vs TT, adjusted risk ratio HR = 1.48, 95% CI: 1.10–2.01, P = 0.010; HR = 1.21, 95% CI: 1.01–1.46, P = 0.038). In the designed genetic model, the dominant genotype TG + GG was associated with a shorter survival time and worse prognosis among lung cancer patients aged less than 60 years (TG + GG vs TT, adjusted risk ratio HR = 1.43, 95% CI: 1.07–1.90, P = 0.014; Table 4).

Table 3

Association between *MMP12* polymorphism rs586701 in genotype models and prognosis of Chinese patients with lung cancer.
Variables	Death/survive			HR(95%Cl)	P	HR^a(95%Cl)	P^a
Variables	TT(ref)		TG
Sex
Male	366/85		123/17	1.23 (1.01–1.52)	0.044	1.24 (1.01–1.52)	0.042
Female	113/43		51/17	1.20 (0.86–1.67)	0.286	1.14 (0.81–1.60)	0.445
Age
≥60	317/62		109/20	1.02 (0.82–1.26)	0.885	1.05 (0.84–1.31)	0.664
<60	162/66		65/14	1.60 (1.19–2.14)	0.002	1.58 (1.18–2.12)	0.002
Smoking stage
Smoker	355/76		117/14	1.30 (1.05–1.60)	0.015	1.30 (1.06–1.61)	0.013
Nonsmoker	116/46		53/18	1.12 (0.81–1.55)	0.512	1.07 (0.77–1.48)	0.706
Family History
Yes	174/43		68/8	1.29 (0.97–1.70)	0.078	1.27 (0.95–1.69)	0.110
No	305/85	106/26		0.99 (0.71–1.39)	0.955	0.99 (0.71–1.39)	0.953
Subtype
ADC	196/64	74/23		1.10 (0.84–1.44)	0.481	1.09 (0.83–1.42)	0.551
SCC	173/38	59/3		1.46 (1.08–1.97)	0.013	1.48 (1.10–2.01)	0.011
NSCLC	434/116	160/33		1.20 (1.00-1.44)	0.046	1.21 (1.01–1.46)	0.038
TNM stage
StageⅠ+Ⅱ	58/59	18/15		1.41 (0.67–2.95)	0.367	1.48 (0.70–3.11)	0.303
Stage Ⅲ+Ⅳ	377/62	149/18		1.08 (0.77–1.51)	0.662	1.04 (0.74–1.46)	0.812

Table 4

Association between MMP12 polymorphism rs586701 in dominant genotype and prognosis of Chinese patients with lung cancer.
Variables	Death/survive			HR(95%Cl)	P	HR^a(95%Cl)	P^a
Variables	TT(ref)		TG + GG
Sex
Male	366/85		136/23	1.15 (0.94–1.40)	0.172	1.15 (0.94–1.40)	0.171
Female	113/43		53/20	1.11 (0.80–1.54)	0.525	1.07 (0.77–1.49)	0.692
Age
≥60	317/62		120/25	0.96 (0.78–1.19)	0.729	0.99 (0.80–1.23)	0.938
<60	162/66		65/14	1.60 (1.19–2.14)	0.002	1.58 (1.18–2.12)	0.002
Smoking stage
Smoker	355/76		132/19	1.21 (0.99–1.48)	0.063	1.21 (0.99–1.49)	0.059
Nonsmoker	116/46		53/22	1.00 (0.72–1.39)	0.982	0.96 (0.69–1.34)	0.813
Family History
Yes	174/43		73/12	1.18 (0.89–1.55)	0.248	1.15 (0.87–1.53)	0.314
No	305/85	116/31		1.09 (0.88–1.35)	0.429	1.10 (0.88–1.36)	0.409
Subtype
ADC	196/64	80/27		1.06 (0.81–1.37)	0.678	1.05 (0.81–1.36)	0.732
SCC	173/38	66/5		1.30 (0.98–1.73)	0.074	1.31 (0.98–1.75)	0.067
NSCLC	434/116	175/42		1.12 (0.94–1.33)	0.222	1.13 (0.94–1.34)	0.189
TNM stage
StageⅠ+Ⅱ	58/59	18/19		1.41 (0.67–2.95)	0.367	1.48 (0.70–3.11)	0.303
Stage Ⅲ+Ⅳ	377/62	163/23		1.04 (0.87–1.25)	0.665	1.04 (0.87–1.26)	0.647

The K-M survival curves demonstrated that the MMP12 polymorphism rs586701 T > G was associated with a reduction in median survival time across a broad spectrum of patients. Kaplan-Meier curves demonstrated that male patients with the TG genotype (MST: 27.37 months) exhibited a shorter MST compared with male patients with the TT genotype (MST: 37.17 M) (log-rank P = 0.046; Fig. 2A). Compared with patients aged < 60 years with the TT genotype (MST: 47.57 M), patients aged < 60 years with the TG genotype (MST: 30.37 M) had a shorter MST (Log-rank P = 0.0016; Fig. 2B), and patients aged < 60 years with the TG + GG genotype (MST: 32.7 M) had a shorter MST (Log-rank P = 0.012; Fig. 2C). The mean survival time of patients with the TG genotype (MST: 25.6 M) was found to be significantly shorter than that of patients with the TT genotype (MST: 37.13 M) (Log-rank P = 0.016; Fig. 2D). SCC patients with the TG genotype (MST: 25.13 M) exhibited a shorter MST compared to SCC patients with the TT genotype (MST: 38.43 M) (Log-rank P = 0.012; Fig. 2E). Patients with non-small cell lung cancer (NSCLC) who had the TG genotype (MST: 30.23 M) exhibited a shorter median survival time (MST) compared to patients with the TT genotype (MST: 39.37 M) (Log-rank P = 0.047; Fig. 2F).

Expression and prognostic associations were analyzed using the Cancer Genome Atlas (TCGA) database. Our findings indicate that the expression of MMP12 polymorphisms is higher in lung cancer tissues (including adenocarcinomas and squamous carcinomas) than in normal tumor walls (Fig. 3A). Furthermore, the expression of MMP12 polymorphisms was found to be higher than that in tumor wall tissues in patients who smoked (Fig. 3B), in patients aged less than 60 years (Fig. 3C), and in male patients (Fig. 3D).

A potential correlation between the MMP12 polymorphism rs586701 and the prognosis of lung cancer was identified through genotyping and follow-up investigation of blood samples from 839 lung cancer patients. n the overall sample, we observed that the rs586701 TG genotype was associated with a worse prognosis in lung cancer. In further stratified analyses, we observed the same results in men, individuals aged < 60 years, smokers, SCC, and NSCLC patients. To the best of our knowledge, our study is the first to investigate the association between the MMP12 polymorphism rs586701 and lung cancer prognosis based on genotype/allele in a Han Chinese population. Our findings indicate that the MMP12 polymorphism rs586701 T>G is associated with a worse prognosis.。

MMPs (matrix metalloproteinases) are a family of proteins structurally and functionally related to zinc endopeptidases that control extracellular protein hydrolysis. They are important regulators of the cellular microenvironment and have been implicated in the invasion and metastasis of a variety of tumor cells. MMP12 expression has been demonstrated to serve as a prognostic marker and therapeutic target in the development of lung cancer in humans and mice,and patients with high MMP12 expression have been shown to have a poorer prognosis [31–33]. One study demonstrated that elevated MMP12 expression was associated with a poor prognosis of lung cancer in mice through a mouse lung cancer model[34]. A study investigating the impact of MMP12 polymorphisms on lung function revealed that MMP12 polymorphisms were associated with reduced lung function in patients with [35]. The present study has identified a significant association between the MMP12 polymorphism rs586701 T>G and poor prognosis in lung cancer. Furthermore, clinical stratification revealed that this association is particularly prevalent in men, individuals aged less than 60 years, smokers, and patients with SCC and NSCLC lung cancer.

The results of our study indicated that the MMP12 polymorphism rs586701T>G was associated with a poor prognosis in male lung cancer patients. The authors concluded that the prognostic differences due to gender factors are mainly closely related to sex hormones and the X chromosome. They further noted that differences in sex hormones between males and females are important biological factors affecting the prognosis of lung cancer[36, 37]. Additionally, they observed that the expression of oncogenes encoded by the X chromosome is higher in female cancer patients[38]. A study conducted on animals and humans revealed that exposure to tobacco pollutants elevated the likelihood of lymphoid aggregation formation and heightened susceptibility to impaired lung function in female mice relative to male mice. Additionally, in human lung tissue, female smokers demonstrated an augmented number of lymphoid follicles and a less favorable prognosis for patients with lung cancer compared to their male counterparts[39]. The aforementioned findings collectively demonstrate that gender is a significant clinical factor that affects the prognosis of lung cancer patients.

A stratified analysis revealed that the MMP12 polymorphism rs586701T > G is associated with a poorer prognosis in patients with smoking-related lung cancer. Macrophages are the primary cell type responsible for the production of MMP-12, a protein that plays a crucial role in the inflammatory response. These cells are the main cell type that normally patrols the lower airways and are the primary inflammatory cell type produced during smoking. Smoking is a major risk factor for lung cancer, and the smoke produced by tobacco combustion contains dozens of lung carcinogens such as aromatic adducts, nicotine, heterocyclic aromatic amines, acrolein, alkyl adducts, tar, etc, which are known to affect gene expression through DNA adduct formation[40], epigenomic modification [41, 42], the function of XPC, a DNA repair protein[43],the inhibition of gluconeogenic synthase kinase 3 ( GSK3) and induces the expression of involucrin (a marker of squamous differentiation)[44],, and regulates the acetylcholine system to affect the proliferation and differentiation of lung cancer cells [45], among other pathways that affect the prognosis of lung cancer. In humans, the expression of MMP12 in alveolar macrophages is approximately ninefold higher in smokers than in nonsmokers[46]. The overexpression of MMP12 in the lower respiratory tract results in elastin degradation, which in turn leads to the formation of elastin fragments. These fragments can initiate a positive feedback loop, further increasing macrophage production in mice and cultured human cells[47]. Furthermore, animal studies have demonstrated that mouse models of lung cancer exposed to secondhand smoke exhibit elevated MMP12 expression (9.3-fold compared to airborne polyurethane controls) and a worse prognosis for lung cancer[34]. The authors of the study concluded that the MMP12 polymorphism rs586701T>G resulted in the upregulation of MMP12 expression, which led to a shorter median survival time and a worse prognosis for the patients.。

The specific type of cancer tissue is also a significant factor influencing the prognosis of lung cancer. In one study, 13 patients with SCC were analyzed with normal lung tissue and 13 patients with adenocarcinoma for related gene expression. It was found that MMP12 expression was significantly up-regulated in patients with SCC [48], while it was also pointed out that MMP12 was negatively correlated with prognosis [49], and that MMP12 deletion could be used as a marker for a good prognosis in SCC[50]. A study that performed a multi-omics analysis of treated non-small cell lung cancer (NSCLC) patients, including multiplex immunofluorescence, nCounter PanCancer Immune Profiling Panel, whole-exome sequencing, and Olink, found that upregulation of MMP12 expression is associated with reduced survival in NSCLC patients[51]. A study demonstrated that atorvastatin significantly reduced the expression of MMP12 in cellular experiments, indicating that MMP12 is a potential target for the treatment of non-small cell lung cancer (NSCLC) patients[52], Furthermore, the lack of MMP12 in lung cancer cells in vivo has been shown to reduce tumor growth and invasiveness [53]. These studies have illustrated that elevated MMP12 expression is associated with a worse prognosis. It is postulated that our findings, which indicate that the MMP12 polymorphism rs586701T>G upregulates MMP12 expression, result in shorter survival times and a poorer prognosis for patients with SCC and NSCLC.

This study is subject to both strengths and limitations. In terms of strengths, we conducted a large-sample study to detect predictive polymorphisms in all primary lung cancer patients. This was done in order to reduce data complexity and prevent important gene deletions. The aim was to investigate the relationship between rs586701 and the prognosis of lung cancer patients. However, it should be noted that the study still has some limitations. Firstly, the study samples were mainly from two hospitals, which may have introduced some selection bias. Secondly, the genetic model assessment was performed retrospectively, which is prone to recall bias. Thirdly, cellular experiments were not performed to investigate the mechanism by which the MMP12 polymorphism rs586701T>G made lung cancer patients' prognosis worse. Additionally, the sample sizes in some of the stratified analyses were relatively small, which may have affected the statistical associations between the tested genotypes and patient characteristics.

This is the inaugural study to investigate the correlation between the MMP12 polymorphism rs586701 and prognosis in lung cancer patients. The study revealed that the MMP12 polymorphism rs586701 was significantly associated with poor prognosis in all lung cancer patients. Furthermore, the study identified that men carrying the rs586701TG genotype, younger than 60 years of age, smokers, and patients with SCC and NSCLC lung cancers all had a poorer prognosis. In addition, the G allele was found to be associated with a poor prognosis in lung cancer patients younger than 60 years of age. Furthermore, both the TG and dominant TG + GG genotypes were also found to have a poor prognosis in lung cancer patients younger than 60 years of age. The relationship between the MMP12 polymorphism rs586701 and the clinical characteristics of specific cancers provides a new target for "individualized anticancer" drug therapy. This can help clinicians to select appropriate individualized patients for target regulation and guide early evidence-based treatment, which is of great value for improving the prognosis of cancer patients.

6 ACKNOWLEDGMENTS

Supported in part through the computational resources and staff expertise provided by Scientific Computing at the Fudan University in Shanghai.

6.1 Authors’ Contributions

X C,W D and ZY S: Writing-original draft. M X,SC G, ZY G and Y W: Supervision; Validation. HJ W, Q L: Visualization.JJ W,B S: Funding acquisition.Y X,GX J,Y B: Writing – review & editing.

6.2 Funding

The work is currently receiving four grants from the National Natural Science Foundation of China (Grant NO. 81372236, 82272863 and 81972822), the Shanghai Postdoctoral Research Foundation in 2021 (Class A Grant NO. 12R21411500).

6.3 Data availability

The data that support the findings of this study are not publicly available because Genomic data is private and cannot be disclosed according to Chinese law.But all data in this study are available upon reasonable request from corresponding author Y B.

6.4 Ethics Approval

The Ethics Committees of the School of Life Sciences of Fudan University (approval number: Discussion and Research Grant No. 244) and Shanghai Changhai Hospital (approval number: CHEC2015-100) have approved this study, and All participants gave informed consent for the study.

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No competing interests reported.

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Influence of MMP12 Single Nucleotide Polymorphism rs586701 on the Prognosis of Primary Lung Cancer in Chinese Han Chinese

Status:

Version 1

Abstract

Purpose

Methods

Results

Conclusion

Figures

1 INTRODUCTION

2 METHODS

2.1 Ethics Statement

2.2 Subjects of study and data collection

2.3 SNP genotyping

2.4 Statistical analysis

3 RESULTS

3.1 Demographic and clinical characteristics and prognostic analysis.

3.2 Correlation between MMP12 polymorphisms and prognosis of lung cancer.

3.3 Stratified analysis of the association between MMP12 polymorphisms and lung cancer prognosis.

4 DISCUSSION

5 CONCLUSION

Declarations

References

Additional Declarations

Status:

Version 1