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Detection of disseminated lung cancer cells in regional lymph nodes by assay of CK19 reverse transcriptase polymerase chain reaction and its clinical significance

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Abstract

Purpose: To set up a molecular method (i.e. RT-PCR) that can be used to detect disseminated tumor cells (DTCs) in regional lymph nodes (LNs) in patients with lung cancer and to evaluate its clinical significance.

Methods: Cytokeratin 19 (CK19) was used as marker. Serial dilution study for LC-5 cells (a lung squamous cell line) was performed to detect sensitivity of the molecular protocol. Regional LNs (n=261) and primary lung cancer tissue (n=40) were obtained from 40 patients with lung cancer who underwent lobectomy or pneumonectomy. They were randomly categorized into two groups: group I (LN-based study, n=20) and group II (patient-based study, n=20). Each LN was halved. One half of a LN was subjected to histological examination (HE) and the other half was subjected to RT-PCR amplification of CK19 mRNA. The effect on survival was analyzed. The cumulative survival was calculated by the Kaplan–Meier method and compared by the log rank test. The Cox model analyzed the prognostic factors.

Results: CK19 mRNA expressed in all tumor tissues as well as LC-5, PAa cells (a lung adenocarcinoma cell line), but not in normal control LNs. Serial dilution study for LC-5 cells demonstrated that CK19 mRNA was detectable at a concentration as low as 10 LC-5 cells in 1×107 LN cells. There was no significant difference between the detecting result of single LN and that of mixed LNs (P>0.05). In 18 of 40 patients, the metastasis in regional LNs was found by both HE and RT-PCR. Of 22 patients without pathologically involved nodes, six (27%) were found to express CK19 mRNA in regional LNs. According to the results of regional LNs in 40 patients by molecular assay, the presence of the CK19 product in LNs was related to tumor size (χ2 =5.76, P<0.025) as well as cell differentiation of the tumor (χ2=7.08, P<0.01). Following a median observation time of 26 months (range, 4 to 60 months), patients with DTCs in nodes showed significant shorter disease-free survival duration than node-negative patients (log-rank test, P=0.001). The independence of this prognostic significance was demonstrated by a multivariate analysis (Cox regression model, P=0.004). The results diagnosed by HE had no significant effect on prognoses (P=0.455).

Conclusions: Comparing with HE, RT-PCR can make more accurate assessment of metastatic status in LNs, which is helpful for screening the patients in whom the early subclinical metastasis exists and disclosing the intrinsic regulation of malignant metastasis. The presence of DTCs in LNs is an independent factor for prognosis. Molecular detection of DTCs in LNs is a supplement for current tumor staging in lung carcinoma.

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Abbreviations

RT-PCR:

Reverse transcriptase-polymerase chain reaction

DTCs:

Disseminated tumor cells

LNs:

Lymph nodes

CK19 :

Cytokeratin 19

HE:

Histological examination

p-status:

The metastatic status of LNs by pathological examination

m-status:

The metastatic status of LNs by molecular method

PAT:

Postoperational adjuvant therapy

LM:

Lymph nodal metastases

SCC:

Squamous cell carcinoma

ADC:

Adenocarcimoma

ASC:

Adenosquamous carcinoma

Mod:

Moderately differentiated

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Authors and Affiliations

  1. Department of Thoracic Surgery [MJ G, QC W and LB L] and Clinical Laboratory [M W and YH Z], The First Affiliated Hospital, Chongqing University of Medical Science, Chongqing, 400016, People’s Republic of China

    Ming Jian Ge, Qing Chen Wu, Mei Wang, Yu Hong Zhang & Liang Bin Li

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  1. Ming Jian Ge
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  2. Qing Chen Wu
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  3. Mei Wang
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  4. Yu Hong Zhang
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  5. Liang Bin Li
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Correspondence to Ming Jian Ge.

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Supported by grant from the Scientific Fund of Chongqing Health Bureau.

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Ge, M.J., Wu, Q.C., Wang, M. et al. Detection of disseminated lung cancer cells in regional lymph nodes by assay of CK19 reverse transcriptase polymerase chain reaction and its clinical significance. J Cancer Res Clin Oncol 131, 662–668 (2005). https://doi.org/10.1007/s00432-005-0009-0

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  • DOI: https://doi.org/10.1007/s00432-005-0009-0

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